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عضویت
فهرست مطالب نویسنده:

gholamreza irajian

  • Narges Nodeh Farahani, Behrooz Sadeghi Kalani, Seyed Hamidreza Monavari, Shiva Mirkalantari, Fatemeh Montazer, Mohammad Sholeh, Zahra Javanmard, Gholamreza Irajian *
    Background and Objectives

    Helicobacter pylori causes several gastrointestinal diseases, including asymptomatic gastritis, chronic peptic ulcer, duodenal ulcer, lymphoma of the mucosa-associated lymphoid tissue (MALT), and gastric adenocarcinoma. In recent years, failure to eradicate H. pylori infections has become an alarming problem for physicians. It is now clear that the current treatment strategies may become ineffective, necessitating the development of innovative antimicrobial compounds as alternative treatments.

    Materials and Methods

    In this experimental study, a cell-penetrating peptide-conjugated peptide nucleic acid (CPP-PNA) was used to target the cagA expression. cagA expression was evaluated using RT-qPCR assay after treatment by the CPP-PNA in cell culture and animal model. Additionally, immunogenicity and toxicity of the CPP-PNA were assessed in both cell culture and animal models.

    Results

    Our analysis showed that cagA mRNA levels reduced in H. pylori-infected HT29 cells after treatment with CPP-PNA in a dose-dependent manner. Also, cagA expression in bacterial RNA extracted from stomach tissue of mice treated with PNA was reduced compared to that of untreated mice. The expression of inflammatory cytokines also decreased in cells and tissue of H. pylori-infected mice after PNA treatment. The tested CPP-PNA showed no significant adverse effects on cell proliferation of cultured cells and no detectable toxicity and immunogenicity were observed in mice.

    Conclusion

    These results suggest the effectiveness of CPP-PNA in targeting CagA for various research and therapeutic purposes, offering a potential antisense therapy against H. pylori infections.

    Keywords: Helicobacter pylori, cag A, Peptide nucleic acid, Immunogenicity, Cytotoxicit
  • Elahe Mirzazade, Gholamreza Irajian, Narges Nodeh Farahani, Abed Zahedi Bialvaei, Farhad Ghasemi Ostad, Bahareh Sadat Rasouli, Amirreza Ehsani, Simin Almasi *
    Background
    The relationship between rheumatoid arthritis and poor oral hygiene has longbeen proven. Colonization of Porphyromonas gingivalis in the oral cavity can play an important role in rheumatoid arthritis. The aim of this study was to determine whether there is a relationship between rheumatoid arthritis disease and the presence of P. gingivalis.
    Methods
    sixty patients with rheumatoid arthritis and 75 controls with matched age, sex and smoking and oral health status were selected. Provided samples from dental plaque were assessed for the presence of P. gingivalis by using bacterial culture and polymerase chain reaction (PCR).
    Results
    Results of bacterial culture and PCR showed no significant difference between rheumatoid arthritis patients and control group in terms of the presence of P. gingivalis (P = 0.9 and P = 0.1, respectively). There was also no relationship between the number of bacterial copies and the activities of rheumatoid arthritis.
    Conclusion
    Findings showed that there was no significant relationship between the presence of P. gingivalis in dental plaque and rheumatoid arthritis.
    Keywords: Rheumatoid Arthritis, Porphyromonas gingivalis, Polymerase Chain Reaction, Periodontitis
  • مریم رضایی، نادیا کاظمی پور، جلیل وندیوسفی*، فرخ رخ بخش زمین، غلامرضا ایراجیان
    زمینه و هدف

    لیستریا منوسیتوژنز یک باکتری درون سلولی اختیاری و عامل بیماری لیستریوزیس است که از طریق مواد غذایی خام و آماده مصرف به انسان منتقل می شود. هدف پژوهش حاضر بررسی سروواریته های غالب لیستریا منوسیتوژنز جداشده از سقط جنین های خودبخودی با روش های فنوتیپی و ژنوتیپی است.

    مواد و روش کار

    دراین مطالعه تعداد 258 نمونه بالینی شامل ترشحات جفت، سواپ واژن و خون از 123 بیمار مبتلا به سقط جنین خودبخودی بررسی شدند و آزمایش های باکتری شناسی، سرولوژی و تسمت بیماریزایی انجام و سروتیپ های غالب با استفاده از Multiplex PCR  شناسایی شدند.

    یافته ها

    در این پژوهش 28 (18/8%) لیستریا منوسیتوژنز جدا گردید که تعداد21 (17/7%)، 5 (5/7%) و 2 (3/7%) از ترشحات جفت، واژینال و خون جدا شدند که 14 (50%) سروواریته a 1/2 و 10 (35/7%) سروواریته b4 و 4 (14/6%) مورد سروتیپ c2، برای نخستین بار از سقط جنین خودبخودی گزارش گردید. 28 مورد (22/76%)  لیستریا منوسیتوژنز با ژن hlyA و 24 مورد با ژن iap  (19/57 %) در مقایسه با گروه شاهد با اختلاف معنی داری مثبت شدند.

    نتیجه گیری

     در این مطالعه مشخص گردید وجود ژن های hlyA و iap افزاینده شدت بیماریزایی بودند و برای اولین بار نشان داده شد سروتیپ های فاقد ژن iap از شدت بیماریزایی کمتری در موش ها برخوردار بودند. این نتیجه حاصل شد که در صورت عدم دسترسی به تشخیص های مولکولی، انجام PI-PLC، کونگورد و بیماریزایی in vivo می توانند در تشخیص لیستریا موثر باشند.

    کلید واژگان: لیستریا منوسیتوژنز، سروتیپ های غالب، ژن های iap و hly، سقط جنین خودبخودی، مولتی پلکس PCR
    Maryam Rezaei, Nadia Kazemipour, Jalil Vandyousefi*, Farokh Rokhbakhshzamin, Gholamreza Irajian
    Background and Aims

    Listeria monocytogenes, a gram positive, facultative, intracellular bacterium is the causative agent of listeriosis that is transmitted to human through raw and ready-to-eat foods. The aim of the present study was to determine dominant serovars of L. monocytogenes isolated from spontaneous abortion using phenotypic and genotypic methods.

    Materials and Methods

    In present study, 258 clinical specimens including placental secretions, vaginal swabs and blood samples from 123 patients with abortion were selected in sterile condition then bacteriological, serological and molecular tests were conducted; dominant serotypes were identified by Multiplex PCR.

    Results

    Out of 28 (%18.8) isolates of L. monocytogenes 21 (%17.7), 5 (%5.7) and 2 (%3.37) were isolated from placental secretions, vaginal swabs and blood respectively. Maximum and minimum isolated of bacteria related to placental secretions and vaginal swabs with 21 and 2 isolates respectively, of which 14 (%50) 1/2a, 10 (%35.7) 4b and 4 (%14.6) 2c serovars were reported for the first time. All of serovars played a key role in the spontaneous abortion as dominant and common serotypes in Iran. All of the isolates 28 (%22.76) showed hlyA gene and 24 isolates (%19.57) were positive for iap gene and compaired with control group there was significant different between the two groups (P<0.0002).

    Conclusion

    The present study showed the isolation dominant and common serotypes of L. monocytogenes from spontaneous abortion and demonstrated that the presence of hlyA and iap were effective genes in increasing aggressive and pathogenicity. Serotypes that lacked the iap gene have less pathogenicity and influenced the pathogenesis in mice. It was also concluded that in the absence of access to molecular tests, performing PI-PLC, Congored and in vivo pathogenicity can be effective in detecting pathogenic serotypes from non-pathogenic L. monocytogenes.

    Keywords: Listeria monocytogenes, Dominant serotypes, iap, hly, Spontaneous abortion, Multiplex-PCR
  • Maryam Rezaei, Nadia Kazemi Pour*, Jalil Vandyousefi, Farokh Rokhbakhsh Zamin, Gholamreza Irajian
    Background and Aims
    The aim was to determine the role of dominant serovars of Listeria monocytogenes (L. monocytogenes) in spontaneous abortions, using isolation methods and polymerase change reaction (PCR).
    Materials and Methods
    A total of 258 samples comprising of placental tissue, vaginal swabs and blood were collected from 123 patients with spontaneous abortion. L. monocytogenes was identified and confirmed by culture, biochemical reactions, serological tests, API system, CAMP (Christie, Atkins, Munch and Petersen) test, and hemolysis on sheep blood agar. Phosphatidyl inositol specific phospholipase C (PI-PLC) assay, followed by multiplex PCR was applied for detection of serotypes 1/2a and 4b.
    Result
    Out of 258 samples, 28 isolates of L. monocytogenes were identified   by different methods. All of the isolates were confirmed by PCR. Of 28 isolated strains, 14(50%) belonged to serovar 1/2a, 10(35.7%) to serovar 4b and 4(14.3%) to other serovars.
    Conclusions
    Based on our study, serovars 1/2a and 4b are dominant serovars as causative agents of human spontaneous abortion due to L. monocytogenes in pregnant women.
    Keywords: L. monocytogenes, Serovars, Spontaneous abortion
  • Mehrdad Gholami, Alireza Salimi Chirani, Reza Falak, Mona Moshiri, Shabnam Razavi *, Gholamreza Irajian*
    Background
    Pseudomonas aeruginosa, an opportunistic pathogen, is a common cause of healthcare-associated infections in immunocompromised individuals. The rapid emergence of multidrug-resistant strains has made P. aeruginosa infections progressively difficult to treat. In this study we evaluated the effect of a chimeric protein containing a P. aeruginosa PilQ fragment and the PilA disulfide loop (PilA-DSL) on the humoral immune response in BALB/c mice.
    Methods
    A chimeric gene encoding an immunogenic region of PilQ and the PilA-DSL was synthesized. Following bacterial expression and purification, the protein was administered to mice and the humoral immune response analyzed. The resulting antibodies were analyzed using an opsonophagocytic killing assay.
    Results
    The anti-recombinant protein antibody titer was significantly greater in immunized mice than in controls. In addition, antibody titers were significantly increased after booster immunizations, and the immunizations induced opsonophagocytosis of P. aeruginosa PAO1.
    Conclusions
    These results suggest that an anti-adhesion-based vaccination may be effective in preventing P. aeruginosa infections. Further studies are needed to evaluate the abilities of such bivalent proteins to induce strong immune responses.
    Keywords: Chimeric protein, PilQ, Pseudomonas aeruginosa, Type IV pili, Vaccine
  • Abed Zahedi Bialvaei, Vajihe Sheikhalizadeh, Ali Mojathedi, Gholamreza Irajian *
    Objective(s)
    Listeria monocytogenes is a foodborne pathogenic bacteria causing the infection listeriosis, which possibly affects all people, particularly immunocompromised persons and pregnant women. This microorganism can be found in several processed foods, dairy products, raw milk, meat and fish products, seafoods, eggs, fruits, and vegetables. This review discusses about the epidemiological significance, incidence, contamination routes of L. monocytogenes in different products and current data about listeriosis in the Iran.
    Materials And Methods
    For accessing to relevant articles and studies, a search was done in main databases and also, almost all Iranian published articles were studied in this field.
    Results
    Outbreaks of listeriosis have been reported in many parts of the worldwide, however there is scanty data about the prevalence of listeriosis in Iran. Accordingly, as a result of high incidence of L. monocytogenes in women with bad obstetric history or history of abortions, diagnosis procedures for detection of L. monocytogenes and timely treatment was suggested.
    Conclusion
    In spite of low incidence of infection in the past, increased interest for lightly preserved and/or ready-to-eat (RTE) food products has recently led to increasing of L. monocytogenes prevalence which has become a public health concern. Subsequently, further researches about the prevalence of L. monocytogenes and also antibiotic susceptibility testing is needed to enable the detection of the contaminated foods, as well as ensures the effective treatment.
    Keywords: Epidemiology, Food, Iran, Listeria monocytogenes, Listeriosis
  • Mehrdad Gholami, Mohammadreza Haghshenas, Mona Moshiri, Shbnam Razavi, Abazar Pournajaf, Gholamreza Irajian *, Mohsen Heidary
    Background and Objective
    Multidrug-resistant Acinetobacter baumannii (MDR-AB) is an important nosocomial pathogen which is associated with significant morbidity and mortality, particularly in high-risk populations. Aminoglycoside-modifying enzymes (AMEs) and 16S ribosomal RNA (16S rRNA) methylation are two important mechanisms of resistance to aminoglycosides. The aim of this study was to determine the prevalence of 16S rRNA methylase (armA, rmtA, rmtB, rmtC, and rmtD), and the AME genes [aac(6′)-Ib, aac(3)-I, ant(3′′)-I, aph(3′)-I and aac(6')-Id], among clinical isolates of A. baumannii in Tehran, Iran.
    Methods
    Between November 2015 to July 2016, a total of 110 clinical strains of A. baumannii were isolated from patients in two teaching hospitals in Tehran, Iran. Antimicrobial susceptibility testing was performed according to Clinical and Laboratory Standards Institute guidelines. The presence of genes encoding the AMEs and16S rRNA methylases responsible for resis­tance was investigated by multiplex polymerase chain reaction.
    Results
    The results showed that colistin was an effective antibiotic and could be used as a last-resort treatment of infections caused by MDR-AB. The resistance rate to aminoglycosides were 100%, 96.36% and 90.9% for tobramycin, gentamicin and amikacin, respectively. In this study, AME genes of aac(6′)-Ib, aac(3)-I and ant(3′′)-I were most prevalent among the isolated strains.
    Conclusion Markedly high resistance to tobramycin, gentamicin and amikacin was noted in current study. Our results suggested that modifying enzyme genes in conjunction with methylation of 16S rRNA might contribute to aminoglycoside resistance induced in vivo in A. baumannii.Further studies are required to determine the prevalence of the aminoglycoside resistance genes in other hospitals of Iran.
    Keywords: Acinetobacter baumannii, Multidrug resistance, Aminoglycoside-modifying enzymes, 16S rRNA methylase
  • Mojtaba Arslani, Tahereh Faramarzi, Maryam Mirzabaygi, Reza Mirnejad, Gholam Reza Irajian, Mojtaba Sade*
    Background
    Foodborne diseases are one of the serious problems in the world. Every year, more than 100 million people are affected by foodborne and waterborne diseases particularly immunocompromised diseases.
    Objectives
    The aim of the present study was to evaluate bacterial load and antibiotic resistance pattern in bacterial isolates from food samples of meat, dairy, and pastry products from west of Tehran, Iran, during April 2007 to March 2008.
    Materials And Methods
    A total of 1625 different food samples including dairy products, meat and pastries were collected randomly from different parts of the west of Tehran. All samples were kept at 4°C. The samples were first cultured according to the standard bacteriological methods and then Staphylococcus aureus and Escherichia coli isolates were identified using standard bacteriological tests. Antimicrobial susceptibility test was performed by disk diffusion method according to Clinical & Laboratory Standards Institute (CLSI) guidelines.
    Results
    During 2007 and 2008, 2.8% and 3% of the food samples were contaminated with S. aureus. Similarly, 3.5% and 6.4% of the food samples were contaminated with E. coli. E. coli isolates were highly resistant to amikacin and cephotaxime and this resistance was increased in 2008. Similarly S. aureus isolates were resistant to ciprofloxacin, cephotaxime, gentamicin, and tetracyclin. There was no significant difference during 2007-2008.
    Conclusion
    The rate of contamination during 2007 was 2.8% and during 2008 was 3% for S. aureus. This strain was isolated from the food samples.
    Further studies should be done to determine the changes of bacterial resistance pattern for various food samples. Thus, the baseline for comparison with future prospective studies should be established, enabling the determination of trends over time.
    Keywords: Antibiotic Resistance, Staphylococcus aureus, Escherichia coli, Food
  • علی احمدی، ملیحه طالبی، الناز پرویزی، غلامرضا ایراجیان*
    زمینه و اهداف
    یکی از مشکلات مهم در جداسازی و تشخیص استرپتوکوکوس پنومونیه از نمونه های بالینی، حساسیت زیاد، اتولیز سریع و درنتیجه ناتوانی در نگهداری ایزوله های پنوموکوک می باشد. ازاین رو برای حفظ ایزوله ها به روش های اختصاصی نگهداری میان مدت و طولانی مدت باکتری موردنیاز است. در این مطالعه کارایی چندین محیط کشت مختلف برای نگهداری ایزوله های استرپتوکوکوس پنومونیه باهم مقایسه و بررسی شد.
    مواد و روش کار
    در سال 1394، دوسویه استاندارد پنوموکوک ATCC 49619 و ATCC 6305،30 ایزوله بالینی، و30 ایزوله فلور نرمال پنوموکوک موجود از مطالعه قبلی انتخاب شدند. سپس برای نگهداری میان مدت، محیط کشت دورسه آگار و برای نگهداری طولانی مدت، سه محیط نگه دارنده شاملSkim milk-Tryptone-Glucose-Glycerin (STGG)، نوترینت براث غنی شده و Todd Hewitt Broth (THB) ساخته شده و کارایی آن ها در نگهداری ایزوله های پنوموکوک ارزیابی شد.
    یافته ها و
    نتیجه گیری
    95 درصد از ایزوله های کشت داده شده روی محیط دورسه پس از 70 روز و80 درصد پس از 4 ماه بعد توانایی رشد داشتند. هر سه محیط نگه دارنده طولانی مدت، تا یک سال کارایی خوبی داشتند، اما نوترینت براث غنی شده توانست تا 2 سال باکیفیت بهتری باکتری را حفظ کند. تفاوتی بین ایزوله های جداشده از بیمار و ایزوله های فلور نرمال در توانایی بقا در محیط وجود نداشت. همچنین مهاجم بودن منشا بالینی ایزوله ها تاثیری روی میزان بقای آن ها نداشت. می توان از محیط کشت دورسه اگار به عنوان یک روش ارزان و ساده و بسیار کاربردی برای نگهداری میان مدت و از محیط نوترینت براث غنی شده برای نگهداری بلندمدت ایزوله های پنوموکوک در آزمایشگاه تشخیص طبی استفاده کرد.
    کلید واژگان: استرپتوکوکوس پنومونیه، نگهداری میان مدت، محیط دورسه، نگهداری بلندمدت
    Ali Ahmadi, Malihe Talebi, Elnaz Parvizi, Gholamreza Irajian*
    Background And Aim
    One of the major problems for the isolation and detection of S. pneumoniae from clinical specimens, high sensitivity, rapid autolysis and thus the inability to hold isolates of pneumococcus. So, it is essential to use specific techniques for the preservation of the organism. In this study several different cultures for S. pneumoniae isolates were compared and evaluated.
    Materials And Methods
    In 2014, 2 ATCC standard , 30 clinical, and 30 normal flora pneumococcal available isolates were selected from a previous study. Dorset egg medium was prepared for midterm preservation, three storage environments, including Skim milk-Tryptone-Glucose-Glycerin (STGG), nutrient broth enriched and Todd Hewitt Broth (THB) was constructed to long-term preservation, and their performance on maintenance pneumococcal isolates was assessed.
    Results and
    Conclusions
    95% of the isolates cultured on Dorset after 70 days and 80% after four months later had the ability to grow. All three long-term storage environment, had a good performance for one year but enriched nutrient broth could keep bacteria with but better-quality up to 2 years. There is no difference between isolates from the patients and isolates from normal flora to survive in the environment. Also aggressive origin of iclinical isolates had no effect on their survival rate. Dorset egg (DE) medium can be considered as a simple, low cost and efficient, medium for midterm preservation and enriched nutrient broth for long term storage of pneumococcal isolates in clinical laboratories.
    Keywords: Streptococcus pneumonia, Midterm preservation, Dorset medium, Long term preservation
  • رسول میرزایی، ملیحه طالبی، فائزه مهدیون، محمد فروزش فرد، غلامرضا ایراجیان*
    زمینه و اهداف
    با افزایش عفونت های ایجادشده توسط استافیلوکوکوس اپیدرمیدیس، افتراق بین سویه های مهاجم و غیر مهاجم و تعیین الگوی مقاومت میکروبی در این سویه ها، ازنظر تشخیص و درمان، دارای اهمیت می باشد.
    مواد و روش کار
    از شهریور 93 تا فروردین 94، تعداد 123 سویه جداشده از بیماران و تجهیزات پزشک از بیمارستان های حضرت رسول (ص) و حضرت علی اصغر(ع) با استفاده از ژن های ica، atlE، agrA، sarA و mecA به روشMultiplex- PCR جهت تمایز سویه های مهاجم و آلوده کننده استفاده شد. همچنین مقاومت به آنتی بیوتیک های سفوکسیتین، لینزولاید، کلیندامایسین، اریترومایسین و تتراسایکلین به روش انتشار در آگار و ونکومایسین به روش E-test، بر اساس استانداردهای (CLSI) بررسی شد.
    یافته ها
    شیوع ژن های ica و mecA به ترتیب در سویه های مهاجم 85% و 75% و در سویه های غیر مهاجم 20% و 51% بود (P<0.05) میزان مقاومت به آنتی بیوتیک های سفوکسیتین، تتراسایکلین، کلیندامایسین و اریترومایسین به ترتیب در سویه های مهاجم: 75%،35%، 37، 76% و در سویه های آلوده کننده: 51%، 40%، 30%، 64% بود. در این مطالعه هیچ گونه مقاومتی نسبت به وانکومایسن و لینزولاید مشاهده نشد.
    نتیجه گیری
    نتایج این مطالعه نشان داد که ژن ica می تواند به عنوان مارکر مناسبی برای تمایز سویه های مهاجم از سویه های آلوده کننده مطرح باشد. همچنین جهت درمان بیماران مقاوم به متی سیلین، ونکومایسین داروی مناسبی می باشد.
    کلید واژگان: استافیلوکوکوس اپیدرمیدیس مهاجم، متی سیلین، دیسک دیفیوژن، Multiplex-PCR
    Rasoul Mirzaei, Malihe Talebi, Faeze Mahdiun, Mohammad Foroozeshfard, Gholamreza Irajian*
    Background And Aim
    With increase Staphylococcus epidermidis infections, discrimination between invasive and non-invasive S. epidermidis strains and determination of drug resistance, for diagnosis and treatment is important.
    Materials And Methods
    From August 2014 to March 2015, 123 strains isolated from patients and medical instruments from Ali Asghar and Hazrat Rasoul hospitals were used to discriminate invasive and contaminating S. epidermidis strains by ica, atlE, agrA, sarA, and mecA genes by Multiplex PCR method. Also, according to the CLSI standards, resistance to cefoxitin, linzeolid, clindamycin, erythromycin and tetracycline were tested by disk diffusion method and vancomycin E-test method.
    Results
    prevalence of ica and mecA genes were 85% and 75% in invasive strains and 20% and 51% in contaminating strains, respectively. The resistance level to cefoxitin, tetracycline, clindamycin and erythromycin antibiotics in invasive strains were: 75℅, 35℅, 37℅, 76℅ and in contaminating strains were; 51℅, 40℅, 30℅, 64℅, respectively. Resistance to vancomycin and linezolid was not observed in this study.
    Conclusions
    This study shown that ica gene can be used as a suitable marker for discrimination between invasive and contaminating isolates. Also, vancomycin is a better choice for the treatment of resistant patients to methicillin.
    Keywords: Invasive Staphylococcus epidermidis, Methicillin, Disk diffusion, Multiplex PCR
  • Shirin Sadr, Davood Darban-Sarokhalil, Gholam Reza Irajian, Abbas Ali Imani Fooladi, Jaleh Moradi, Mohammad Mehdi Feizabadi
    Background And Objectives
    For further confirmation of the previous in-house real-time PCR and CYP 141 target as a rapid and cheap diagnostic technique and a new target for direct detection of Mycobacterium tuberculosis, we evaluated and compared the results of smear, culture and real-time PCR in sputa that were collected from 2 health centers. Moreover we tried to evaluate the diagnostic accuracy of phenotypical methods for detection of tuberculosis that have been applied in two health centers of Iran.
    Materials And Methods
    Thirty two sputa (including 15 smear positive and 17 smear negative) and 53 Sputa (29 smear and culture positive and 24 smear and culture negative specimens) were collected from tuberculosis suspected patients from health center No. 1 and 2 respectively. A Taqman probe was used for direct detection of M. tuberculosis using the specific primers.
    Results
    Because of the results, data of health center No. 1 was not reliable. The average number of bacteria that was detected in health center No. 2 by real time PCR was 1.2E퍍7.3퍎; 1.4E퍎1.29퍎 and 8.27E퍎1.07퍎 for one to three plus smear result groups, respectively. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of real-time PCR were 96.5% (28/29), 95.8% (23/24), (96.6%) and (96%), respectively.
    Conclusion
    Compared with the results of previous studies and being a good correlation between real-time PCR and phenotypic methods, emphasize that CYP141 is a good target for quantification of M. tuberculosis in sputa and real-time PCR can be a good method for evaluation of smear microscopy. Moreover, further surveillance is needed to evaluate the phenotypical methods and final decisions that are taken in health centers of Iran that can be observed and evaluated by the cheap molecular methods like in-house methods.
    Keywords: Mycobacterium tuberculosis, Real-time PCR, Cytochrome P450 cyp141
  • فرحناز بینشیان، غلامرضا ایراجیان، محمدحسن باقری منصوری، زهره شریفی*
    زمینه و اهداف
    اینترلوکین10 سایتوکین ضد التهابی مهمی است وپلی مورفیسم های تک نوکلئوتیدی در پروموتر ژن اینترلوکین10 با عفونت Hepatitis B virus))HBV مرتبط بوده بخصوص پلی مورفیسم تک نوکلئوتیدی در منطقه پروموتر ژن10 - IL ، در موقعیت 1082-،819-،592- با تولید 10- IL مرتبط است. در مطالعه حاضر ارتباط بین پلی مورفیسم 819-،10- IL با عفونتHBV مزمن در بیماران ایرانی مورد مطالعه قرارگرفت.
    مواد و روش کار
    در این مطالعه تعداد 200 نفر در دو گروه(100نفرمبتلا به هپاتیت مزمن،100نفردهنده خون)، مبتلا به عفونت HBV مزمن و اهدا کننده خون به صورت تصادفی با محدوده سنی 59-18 سال وارد مطالعه شدند. مطالعه در سال1392بر روی نمونه های بیماران مراجعه کننده به آزمایشگاه تشخیص بالینی انتقال خون انجام شد . استخراج DNA ژنومی از لایه بافی کوت به روش نمک اشباع انجام گردید. از روش PCR اختصاصی آلل (Allele Specific Polymerase Chain Reaction Method) برای تعیین ژنوتیپ های پلی مورفیسم (C/T) 819- پروموتر ژن 10IL- استفاده شد. سپس برای شناسایی محصول PCR، الکتروفورز بر روی ژل آگارز 5/1 درصد انجام گردید. آنالیز اطلاعات به کمک آزمون Chi Square مورد بررسی قرارگرفت.
    یافته ها
    فراوانی ژنوتیپ های هموزیگوت (TT، TC، CC) در موقعیت 819 − ،10- IL در بیماران مبتلا به عفونت HBV مزمن به ترتیب 84و12، 4 درصد و در افراد اهدا کننده خون 89 و10، 1 درصد بود و تفاوت معنی داری بین دو گروه از نظر ژنوتیپی مشاهده نشد (8/0P=). فراوانی آلل واریانت C درگروه بیماران مبتلا به HBV 10 درصد و در افراد سالم 6 درصد بود که از نظر آماری تفاوت معنی داری بین دو گروه مشاهده نگردید (6/0P=).
    نتیجه گیری
    در این مطالعه ارتباط معنی داری بین پلی مورفیسم −819C/T ، 10- IL در دو گروه کنترل و بیماران مبتلا به عفونتHBV مزمن مشاهده نگردید.
    کلید واژگان: اینترلوکین10، هپاتیت B، پلی مورفیسم
    Farahnaz Bineshian, Gholamreza Irajian, Mohammad Hasan Bagheri Mansoori, Zohre Sharifi*
    Background And Aim
    Anti-inflammatory cytokine IL-10 is important and single nucleotide polymorphisms in the interleukin-10 gene promoter is associated with HBV infection. Especially single nucleotide polymorphisms in the promoter region of the gene IL10, at position -1082, -819, -592 is associated with production of IL10. Therefore, the present study was to determine the effect of IL-10 -819 polymorphism with chronic HBV infection in Iranian patients.
    Materials And Methods
    A total of 200 subjects (100 chronically infected hepatitis B patients and 100 blood donors) with an age range of 18-59 years were included in this study randomly.The study in 1392 on samples of patients referred to clinical laboratory blood transfusion was performed. Genomic DNA was extracted from Buffy coat layer with using of salting out procedure.Allele Specific Polymerase Chain Reaction Method was used for determining polymorphisms -819 (C/T) in the IL-10 gene promoter.Electrophoresis was performed on a arose gel 1.5 per cent for the detection of PCR products. Data were analyzed using Chi Square analysis
    Results
    The frequency of Homozygote genotype IL-10 -819 (TT, TC, CC) was in patients with HBV 84%,12% and 4% and 89%,10% and 1% in healthy individuals respectively and no significant difference was observed between the two groups in terms of genotype (P =0.8). Variant of C allele frequency was in patients with HBV 10% and 6% in healthy individuals. There was no statistically significant difference between the two groups (P=.0.6).
    Conclusions
    Our results showed that there was no correlation between polymorphism IL-10-819(T/C) in control group and chronic Hepatitis B infection.
    Keywords: Interleukin-10_chronic Hepatitis B infection_polymorphism
  • شیرین شهسواری*، سیاوش سلمانزاده، اهرابی طاهره فلسفی، غلامرضا ایراجیان
    زمینه و اهداف
    با توجه به رشد روبه افزایش باکتری های تولید کننده ESBL، در میان اعضای خانواده Enterobacteriaceae، ردیابی و درمان عفونت های بیمارستانی ناشی از ایزوله های بالینی تولید کنندگان ESBL متعلق به این خانواده، دارای اهمیت است. هدف از این پژوهش، ارزیابی یک روش سریع، با استفاده از محیط کلریمتریک جدید ساخته شده در دانشگاه الزهراء (تهران، ایران)، می باشد.
    مواد و روش کار
    ایزوله های بالینی جدا شده از بیماران متعلق به خانواده انتروباکتریاسه، برای ردیابیESBLs با استفاده از دیسک های ceftazidim (CAZ)، ceftazidime + clavulanate (CZA)، cefotaxime +clavulanate (CTC) وCefotaxime (CTX) در دو محیط مولر- هینتون آگار و محیط کلریمتریک در این مطالعه، مورد ارزیابی قرار گرفتند.
    یافته ها و
    نتیجه گیری
    در محیط کلریمتریک، در مدت 6-5 ساعت، به دلیل فعالیت متابولیکی باکتری، تغییر رنگی در محیط کشت رخ داده و محیط از رنگ قرمز به زرد تغییر کرده و در اطراف دیسک ها هاله های مهاری قرمز رنگ ایجاد می گردد. نتایج به دست آمده با روش کلریمتریک مشابه با نتایج به دست آمده از روش استاندارد انتشار دیسک در مولر- هینتون آگار پس از یک شبانه روز، می باشد.
    کلید واژگان: محیط کشت کلریمتریک، ESBLs، تشخیص سریع، آزمون انتشار دیسک
    Shirin Shahsavari*, Siavosh Salmanzade Ahrabi, Tahereh Falsafi, Gholam Reza Irajian
    Background And Aim
    Due to the increased growth of bacteria producing ESBL, among the members of the Enterobacteriaceae family, monitoring and treatment of nosocomial infections are important, especially caused by ESBL-producing clinical isolates belonging to this family. The aim of this study is to evaluate a rapid method, using new colorimetric culture was made at the University of Al-Zahra (Tehran, Iran).The used method to detect ESBLs, according to disk diffusion method based on CLSI on colorimetric culture and the results of it, compared with the Müller-Hinton agar for disk diffusion method.
    Materials And Methods
    In this study, the clinically isolated samples belonged to the Enterobacteriaceae family which was evaluated in both Müller-Hinton agar medium and colorimetric culture using ceftazidime (CAZ), ceftazidime clavulanate (CZA), cefotaxime clavulanate (CTC) and Cefotaxime (CTX) discs.
    Results &
    Conclusions
    Because of the metabolic activity of bacteria, a color change occurred from red to yellow in colorimetric culture after 5-6 hours and red inhibitory zone created around the disc. The obtained results from the colorimetric method are same with the results of Müller-Hinton agar method for disk diffusion after a day.
    Keywords: Colorimetric medium, ESBLs, Rapid detections, Disk diffusion test
  • Faramarz Masjedian Jazi, Gholamreza Irajian*, Reza Mirnejad, Vahhab Piranfar, Taghi Zahraei Salehi, Noor Amir Mozafari, Ehsanollah Ghaznavi, Rad, Mahmoud Khormali
    Background
    Brucellosis is an endemic zoonotic disease in the Middle East. This study intended to design a uniplex PCR assay for the detection and differentiation of Brucella at the species level and determining the antibiotic susceptibility pattern of Brucella in Iran.
    Methods
    Sixty-eight Brucella specimens (38 animal and 30 human specimens) were analyzed using PCR (using one pair of primers). Antibiotic susceptibility patterns were evaluated and compared using the E-Test and disk diffusion susceptibility test. Tigecycline susceptibility pattern was compared with other antibiotics.
    Results
    Thirty six isolates of B. melitensis, 2 isolates of B. abortus and 1 isolate of B. suis from the 38 animal specimens, 24 isolates of B. melitensis and 6 isolates of B. abortus from the 30 human specimens were differentiated. The MIC50 values of doxycycline for human and animal specimens were 125 and 10 μg/ml, respectively, tigecycline 0.064 μg/ml for human specimens and 0.125μg/ml for animal specimens, and trimethoprim/ sulfamethoxazole and ciprofloxacin 0.065 and 0.125μg/ml, respectively, for both human and animal specimens. The highest MIC50 value of streptomycin in the human specimens was 0.5μg/ml and 1μg/ml for the animal specimens. The greatest resistance shown was to tetracycline and gentamicin, respectively.
    Conclusion
    Uniplex PCR for the detection and differentiation of Brucella at the strain level is faster and less expensive than multiplex PCR, and the antibiotics doxycycline, rifampin, trimethoprim-sulfamethoxazole, ciprofloxacin, and ofloxacin are the most effective antibiotics for treating brucellosis. Resistance to tigecycline is increasing, and we recommend that it be used in a combination regimen.
    Keywords: Uniplex PCR, Brucella, Antibiotic Susceptibilities, Tigecycline
  • Farzaneh Khodaei, Ali Ahmadi, Shirin Sayahfar, Gholamreza Irajian, Malihe Talebi*
    Background
    Pili in Streptococcus pneumoniae have been shown to be one of the adherence factors for epithelial cells in the human upper respiratory tract. Two types of pilus-like structures (pilus islet-1 and pilus islet-2) have been distinguished in S. pneumoniae.
    Objectives
    To investigate the presence of pilus islet-1 (PI-1) in S. pneumoniae and the correlation between our isolates.
    Materials And Methods
    In this study, 162 S. pneumoniae isolates were collected from clinical specimens, and normal flora were also examined for the distribution of PI-1 using the presence of the rlrA and rrgC genes as markers for this islet and sipA as an indicator of pilus islet-2 (PI-2). BOX-PCR analyses were performed to determine the genetic relationship between isolates.
    Results
    The results confirmed the presence of rlrA and rrgC genes in both clinical (n = 39) and normal flora (n = 26) isolates. The minimal inhibitory concentration results revealed that the rate of resistance of these isolates to the three antibiotics tested ranged from 26% for penicillin to 46% for erythromycin and tetracycline. Furthermore, 12% of the isolates were resistant to all three antibiotics. Strain typing using repetitive element BOX-PCR analysis among the 65 isolates identified 8 different band patterns.
    Conclusions
    Our results indicated that the dissemination of PI-1 was widespread in S. pneumoniae isolates, although no PI-2 isolates were detected. Furthermore, the frequency of rlrA and rrgC of clinical isolates was significantly more than that of normal flora isolates.
    Keywords: Pilus, BOX, PCR, Streptococcus pneumonia
  • Gholamreza Irajian, Mehri Sharifi, Shiva Mirkalantari, Reza Mirnejad, Mohammad Reza Jalali Nadoushan
    Background
    In most cases, prostatitis can be caused by a bacterial agent such as Ureaplasma urealyticum. Considering to the cumbersome of the culture method for the detection of Ureaplasma species in clinical samples such as prostate; PCR method that is faster and more appropriate than the cultivation methods, can be utilized for the detection of U. urealyticum and U. parvum. PCR-RFLP method can differentiate both biovars and assist in studies of the clinical diagnosis, epidemiology and pathology of this species in human. The aim of this study was to molecular detection of U. urealyticumin in prostate tissue samples based on PCR- RFLP.
    Methods
    Two hundred prostate tissue samples were collected from patient suffering from prostatitis. The PCR assay was used to amplify a 559 bp fragment of 16S-23SRNA interspace region of Ureaplasma. After sequencing, PCR products from positive samples were digested with TaqI restriction enzyme.
    Results
    Seven cases (3.5%) out of 200 prostate tissue samples were positive for U. urealyticum. Results of PCR products sequencing demonstrated that all isolates were U. parvum biovar. PCR-RFLP results shown that there was not any differentiation in pattern of enzymatic digestion, in addition, all isolates were U. parvum, serovar 3.
    Conclusion
    U. urealyticum can be one of the causing agents of prostatitis. Using PCR-RFLP with specific primer and restriction enzyme is a rapid and cost-effect method for detection and differentiation of Ureaplasma from clinical samples.
    Keywords: Ureaplasma urealyticum, Ureaplasma parvum, PCR, RFLP, Prostatitis, Prostate tissue
  • فاطمه کرپی، بهادر بهروز، محمد معتمدی فر، غلامرضا ایراجیان*
    زمینه و اهداف
    سویه های پاتوژن سودوموناس آئروژینوزا پیلی قطبی تولید می کند که برای تحرک، چسبندگی و تهاجم مورد نیاز است. هدف از این مطالعه تولید و تخلیص پروتئین نوترکیب پیلین می باشد.
    مواد و روش ها
    ژن کد کننده پیلین (pilA) از سویه PAO1 سودوموناس آئروژینوزا با بوسیله PCR جدا گردید و در وکتور بیانی pET-22b کلون شد. تایید ساختار وکتور نوترکیب به وسیله ی هضم آنزیمی دوگانه و تعیین توالی صورت گرفت. وکتور نوترکیب به داخلی باکتری (E.coli BL21(DE3 ترانسفورم شد و تخلیص پروتئین نوترکیب بوسیله کروماتوگرافی تمایلی انجام شد. آزمون وسترن بلات با استفاده از توسط آنتی بادی ضد پلی هیستیدنی انجام شد.
    یافته ها
    نتایج تعیین توالی و هضم آنزیمی صحت کلونینگ را تایید نمود. الکتروفورز پروتئین نشان داد که وزن مولکولی پروتئین نوترکیب پیلین حدود 19 کیلودالتون می باشد. همچنین نتایج آزمون وسترن بلات تولید پروتین نوترکیب را تایید کرد. مقدار پروتئین تولید شده که به روش اسپکتروفتومتری مستقیم اندازه گیری شد که مقدار ان.58/2 میلی گرم به هر میلی لیتر تعیین شد.
    نتیجه گیری
    نتایج وسترن بلات و الیزا به موازات نتایج تعیین توالی نشان دهنده صحت تولید پروتئین نوترکیب پیلین و حفظ ساختار نسبی آن می باشد.
    کلید واژگان: پروتئین پیلین سودوموناس آئروژینوزا، کلونینگ، بیان
    Fatemeh Korpi, Bahador Behrouz, Mohmmad Motameifar, Gholamreza Irajian*
    Background
    Pathogenic Pseudomonas aeruginosa strains produce polar pili has required for motility, adhesion, and invasion. The main aims of the present study are to identify, clone, express and purify the recombinant pilin protein of P. aeruginosa in the prokaryotic host.
    Material and
    Methods
    The recombinant pilin gene (pilA) was isolated from P. aeruginosa PAO1 strain by PCR and cloned into pET-22b vector. The recombinant plasmid was subsequently verified by restriction analysis, and DNA sequencing. The recombinant vector was transformed into E. coli BL21 (DE3) strain, then the recombinant pilin overexpressed and affinity purified by Ni-NTA agarose-affinity chromatography.Western blot analysis was performed using anti-6His tag antibody.
    Results
    The PCR and enzymatic digestion results showed the accuracy of the pilA gene cloning. The protein electrophoresis showed that the molecular weight of recombinant pilin is about 19 kDa. Western blot analysis also confirmed the production of recombinant protein. The amount of produced protein was measured by the direct spectrophotometery method, which was 2/58 mg/mL.
    Conclusion
    Western blot and ELISA results along with that of sequencing ensure accurate production of recombinant pilin, retaining its partial epitopes.
    Keywords: Pseudomonas aeruginosa pilin protein, Cloning, Expression
  • Mahdieh Neghabi, Hajiagha, Atousa Aliahmadi, Mohammad Reza Taheri, Alireza Ghassempour, Gholamreza Irajian, Hassan Rezadoost, Mohammad Mehdi Feizabadi
    Background And Objectives
    Due to the importance of finding of new antibacterial agents, the antibacterial properties of Prosopis cineraria aerial parts were investigated using a bioassay guided fractionation scheme.
    Materials And Methods
    The organic extract was prepared via maceration in methanol, followed by the fractionation using n-hexane and ethyl acetate. The MICs of fractions were determined against some human pathogenic bacteria using broth micro-dilution assay. The primary characterization and identification of bioactive substance(s) was based on a bio-autograph- ical method using HPTLC and flash chromatography in parallel with agar overlay assays. Finally the exact mass of effective compound(s) was determined by LC-MS.
    Results
    The best antibacterial activities were related to the ethyl acetate fraction. The effective antibacterial compound of the plant were 2 substances with molecular weight of 348 and 184 Dalton that inhibited the growth of assessed Gram positive bacteria with MIC values lower than 125 to 62.5 µg/ml synergistically.
    Conclusion
    Further analysis using nuclear magnetic resonance could reveal the exact structure of these two antibacterial substances. These 2 effective antibacterial compounds could be applied as lead compound for synthesis of new antibacterial agents.
    Keywords: Staphylococcus aureus, Prosopis cineraria, Antibiotic resistant, characterization, bioassay
  • Abbas Bahador, Behrooz Sadeghi Kalani, Firuzeh Valian, Gholamreza Irajian *, Lida Lotfollahi *
    Background
    Listeria monocytogenes is a foodborne pathogen and a serious threat to the public health in the world. Consumption of traditional foods such as dairy and meat products can be a major reason for relative abundance and isolation of these bacteria..
    Objectives
    The purpose of this study was to determine the phenotypic and genotypic characteristics of L. monocytogenes strains isolated from dairy and meat products..
    Materials And Methods
    A total of 317 dairy products and meat-processed samples were collected. Antibiotic susceptibility test was performed on each sample by the disk diffusion method (Kirby Bauer). Five reference loci were used for typing of L. monocytogenes strains by MLVA (Multiple Locus VNTR Analysis) Technique..
    Results
    A total of 24 L. monocytogenes isolates were collected from the dairy and meat products. Resistance of isolated L. monocytogenes strains to penicillin G were 54.54% (from dairy products) and 46.15% (from processed meat). Genetic relatedness of isolates were assessed by MLVA. Out of 13 different types, type 2 with 6 strains and type 3 with 4 strains, were the most common types..
    Conclusions
    MLVA analysis showed that samples obtained from different sources could have similar genetic profile. As a result, administration of penicillin in patients with listeriosis (especially pregnant women) and antibiotic susceptibility test are recommended. The fast and accurate methods such as MLVA for tracking of pollution sources of L. monocytogenes are recommended during outbreaks..
    Keywords: Genotypic, Dairy Products, Meat, Listeria monocytogenes
  • Gholamreza Irajian, Farahnaz Bineshian, Shokrollah Mashoori, Mohammadreza Freidounian
    Background
    Dermatomycoses are a major public health problem. Tinea cruris is a form of Dermatomycoses.Tinea cruris is a common infection in the groin area, genitals, pubic area, perineum and perianal skin.The purpose of thisstudywas to investigate the cutaneous fungal infections among students in a school dormitory.
    Materials And Methods
    The study was performed on 110 male students dormitory aged between 18 to 28 years.From the specimens obtained, direct preparations were made using %10 potassium hydroxide and cultured on sabouraud dextrose agar containing 0.05 mg/ml chloramphenicoland 0.5 mg/ml cycloheximide (Scc). Identification of fungi was performed according to morphologic and microscopic growth of the colonies and using slide culture method.
    Results
    17 people out of 110 individuals were diagnosed with cutaneous fungal infections. Six people were diagnosed with dermatophytosis. All cases of dermatophytosis were Tinea cruris. Epidermophyton floccosum, the anthropophilic species, was only isolated dermatophyte.The prevalence of Tinea cruris was 5.5% and the Prevalence of Pityriasis versicolor was 4.5%.
    Conclusion
    The most important source of transmission of anthropophilic dermatophyte species was by men to men.The results of this study is similar to most other studies in Iran and other countries. In addition to treatment, other necessary steps should be applied in order to prevent infection and reduce the risk of pathogens.
    Keywords: Dermatomycoses, Dermatophytosis, Epidermophyton, Tinea cruris
  • Ali Ahmadi, Farzaneh Khodaei, Maryam Esghaei, Gholamreza Irajian, Malihe Talebi
    Introduction
    Streptococcus pneumoniae (S. pneumoniae) cause morbidity and mortality in infants and younger children. Because of high prevalence of penicillin resistance, rapid and reliable diagnostic techniques for penicillin non-susceptible S. pneumoniae (PNSSP) are important for prevention and treatment. We investigated the association of the restriction length polymorphism (RFLP) patterns for pbp2b to distinguish between penicillin susceptible and resistant S. pneumoniae isolates.
    Methods
    In this study, a total of 70 pneumococcal isolates were collected from different clinical sources. MIC of these isolates was determined and pbp2b gene was amplified by PCR and they were digested by HaeІІІ enzyme.
    Results
    Of the 70 isolates, 86% (60) and 14% (10) pneumococcal isolates were found to be PNSSP (penicillin intermediate S. pneumoniae (PISP) and penicillin resistant S. pneumoniae (PRSP)) and penicillin susceptible S. pneumoniae (PSSP). In addition, 10 RFLP patterns (A-J)which were based on the HaeІІІ digestion of pbp2b gene were observed. All PSSP isolates showed that they belonged to pattern D, whereas, all PNSSP showed 10 different patterns.
    Conclusion
    In general, the present study suggests that RFLP can be a powerful tool in differentiation between the penicillin resistant and susceptible strains.
  • Elnaz Parvizi, Alireza Nateghian, Ali Ahmadi, Kiana Mirsaeedi, Gholamreza Irajian*
    Streptococcus pyogenes is one of the most important causes of bacterial pharyngitis. Asymptomatic carriage of this organism especially among schoolchildren is a common issue. Study of the prevalence and antimicrobial susceptibility pattern of the flora strains, as clinical indicators, are useful for treatment of streptococcal infections. The aim of this study was to determine the prevalence and resistance pattern of S. pyogenes isolates detected from throat of healthy children in Tehran. After filling a questionnaire including general information, from preschoolers, primary school students and school age children referring to the follow up center of the Ali-Asghar hospital of pediatrics, throat samples were collected from 5-15 year old eligible children by a sterile cotton swab. Then the samples were immediately seeded onto %5 sheep blood agar media. The plates were streaked and incubated appropriately after transferring to the laboratory. Biochemical and serological identification of isolates were done and then Antimicrobial susceptibility pattern of all identified isolates was determined by the disk diffusion method. Finally, Penicillin, Erythromycin and clindamycin MICs were determined by the E-test method for all isolates.The total number of 423 sample swabs were collected during a period of 6 months, showing the carriage rate of %5.7(n=24).Using chisquare test showed that there were no significant differences in carriage rates between the age sub groups (p=0.095). All isolates were sensitive to Penicillin, Cefotaxime, Erythromycin, Vancomycin, Azithromycin and Clindamycin. The rate of intermediate sensitivity and resistance to Tetracycline was 25 and 12.5%, respectively. Two isolates had intermediate sensitivity to each of the agents Oflxacin and Chloramphenicol. The MIC level of Penicillin for all isolates were ≤0.016 μg/ml, and MIC level of clindamycin and erythromycin for all isolates were ≤0.25 μg/ml, which were in sensitive range.It is concluded that in contrast to thepublished reports about rising penicillin MIC and resistance to erythromycin, Penicillin remains the first choice of drug for streptococcal infections and also macrolides and lincosamides can be considered as the alternative choice of drug in allergic patients.
    Keywords: Streptococcus pyogenes, healthy children, Antibiotic susceptibility, MIC
  • Mehrdad Gholami, Alireza Salimi Chirani*, Mona Moshiri, Mansour Sedighi, Abazar Pournajaf, Masoud Tohidfar, Gholamreza Irajian
    Background
    Neisseria meningitidis, a life-threatening human pathogen with the potential to cause large epidemics, can be isolated from the nasopharynx of 5–15% of adults. The aim of the current study was to evaluate biophysical and biochemical properties and immunological aspects of chimeric acyl-carrier protein-macrophage infectivity potentiator protein-type IV pilus biogenesis protein antigen (ACP-MIP-PilQ) from N. meningitidis serogroup B strain.
    Methods
    Biochemical properties and multiple alignments were predicted by appropriate web servers. Secondary molecular structures were predicted based on Chou and Fasman, Garnier-Osguthorpe-Robson, and Neural Network methods. Tertiary modeling elucidated conformational properties of the chimeric protein. Proteasome cleavage and transporter associated with antigen processing (TAP) binding sites, and T- and B-cell antigenic epitopes, were predicted using bioinformatic web servers.
    Results
    Based on our in silico and immunoinformatics analyses, the ACP-MIP-PilQ protein (AMP) can induce high-level cross-strain bactericidal activity. In addition, several immune proteasomal cleavage sites were detected. The 22 epitopes associated with MHC class I and class II (DR) alleles were confirmed in the AMP. Thirty linear B-cell epitopes as antigenic regions were predicted from the full-length protein.
    Conclusion
    All predicted properties of the AMP indicate it could be a good candidate for further immunological in vitro and in vivo studies.
    Keywords: Chimeric protein, In silico, Neisseria meningitides, serogroup B, Vaccine
  • علی احمدی، ملیحه طالبی، شیرین سیاح فر، غلامرضا ایراجیان*
    سابقه و هدف
    استرپتوکوکوس پنومونیه یکی از مهم ترین پاتوژن های باکتریائی و عضوی از استرپتوکوک های ویریدانس بشمار می رود. تشخیص صحیح و تمایز این باکتری از سایر باکتری های مشابه، اهمیت بسیاری در اپیدمیولوژی بالینی و روند مقاومت های آنتی بیوتیکی پنوموکوک دارد. در این مطالعه میزان صحت تشخیص استرپتوکوک پنومونیه در آزمایشگاه های تشخیص طبی با روش های فنوتایپی و ژنوتایپی مورد بررسی قرار گرفت.
    مواد و روش ها
    ایزوله های بالینی که توسط آزمایشگاه های تشخیص طبی به عنوان پنوموکوک شناسایی شده بودند، دریافت و به آزمایشگاه منتقل شد. پس از خالص سازی کلنی ها، تعیین هویت بیوشیمیایی با استفاده از آزمون های حساسیت به اپتوچین و حلالیت در صفرا انجام گردید. DNA ایزوله ها با استفاده از کیت استخراج شده و تعیین هویت مولکولی از طریق آزمون PCR برای تشخیص ژن lytA انجام شد.
    یافته ها
    پس از تعیین هویت مجدد، در نهایت 50 ایزوله به عنوان پنوموکوک واقعی شناسایی شده و 60 ایزوله دیگر شامل 3 کوکوباسیل گرم منفی، 7 استرپتوکوک غیرآلفاهمولیتیک و 50 استرپتوکوک ویریدانس بودند. بیش ترین میزان اشتباه در تشخیص پنوموکوک، مربوط به عفونت های تنفسی و چشمی بود. تمام 50 ایزوله که به عنوان پنوموکوک شناسایی شده بودند، از نظر ژن lytA مثبت بوده و ایزوله های مقاوم به اپتوچین از نظر ژن lytA منفی بودند.
    نتیجه گیری
    در این مطالعه، میزان خطا در تشخیص پنوموکوک 55 درصد تعیین شد. به نظر می رسد استفاده از دیسک های اپتوچین به تنهایی و عدم انجام آزمایشات تکمیلی و فقدان کنترل کیفی مناسب، علل اصلی عدم تشخیص درست پنوموکوک در ایران است. تشخیص غلط باعث اطلاعات اپیدمیولوژیک نادرست، درمان های غیرضروری و افزایش مقاومت آنتی بیوتیکی در باکتری می شود. رعایت مسائل فوق الذکر و حضور یک متخصص میکروب شناسی در آزمایشگاه بیمارستان می تواند بسیار موثر باشد.
    کلید واژگان: استرپتوکوک پنومونی، واکنش زنجیره ای پلیمراز، فنون تشخیصی مولکولی
    Ali Ahmadi, Malihe Talebi, Shirin Sayah Far, Gholamreza Irajian*
    Introduction
    Streptococcus pneumoniae, is one of the most important bacterial pathogens and a member of viridians streptococci group. Accurate identification and differentiation of this form of bacteria from other relative streptococci, is the base of epidemiological study of this type of organism. The aim of this study was to evaluate the accuracy of detection of Streptococcus pneumoniae in clinical laboratories in Tehran, by using phenotypic and genotypic methods.
    Materials And Methods
    A total number of 110 isolates, identified as pneumococci by some clinical laboratories in Tehran, were collected between March 2010 to May 2012.After isolating the colonies, biochemical identification tests by optochin susceptibility (Mast) and bile solubility (direct method) methods were performed. After DNA extraction, PCR was performed to define lytA gene as a molecular identification for Streptococcus pneumonia.
    Results
    After re-identifying the isolates, fifty of them were determined as true pneumococci, and other remaining sixty isolates were identified as: three gram negative coccobacilli, seven non alpha hemolytic streptococci, and fifty Viridans streptococci. Most of misidentifications were related to respiratory and eye infecting streptococci. Unlike non pneumococcal isolates, all 50 pneumococcal isolates were positive for lytA gene.
    Conclusion
    There was 55% error in detection of pneumococci in this study. The use of optochin susceptibility test as the sole detection tool and also lack of supplemental tests and proper quality controlling, are the main causes of failure in diagnosing pneumococci in Iran. Misidentifications may result in incorrect epidemiological data gathering, unnecessary treatment, and false increased antibiotic resistance reports for this organism. Regarding the high incidence of inaccuracies in defining this specific type of microorganism, we suggest the presence of a clinical microbiologist in the hospital laboratories to perform the right diagnostic tests and quality controlling would be essential.
    Keywords: Streptococcus pneumonia, Polymerase chain reaction, Molecular diagnostic techniques
  • Masoume Bina, Abazar Pournajaf, Shiva Mirkalantari, Malihe Talebi, Gholamreza Irajian
    Background And Objective
    The production of carbapenemases especially Klebsiella pneumoniae carbapenemase (KPC) is the most important mechanism of enzymatic resistance in isolated Enterobacteriaceae such as K. pneumoniae. The purpose of this study was detected of the carbapenemase producer K. pneumoniae strains with phenotypic and genotypic methods.
    Method
    Out of 800 strains, 270 K. pneumoniae strains (33.7%), were obtained. Antibiotic susceptibility test was performed by disk diffusion method in accordance with CLSI guidelines. Carbapenem resistant strains were identified by the Modified Hodge Test based on CLSI instruction and PCR for surveying the presence of bla-KPC gene.
    Results
    A total 270 K. pneumoniae strains were collected. Antibiotic susceptibility test results showed the highest and lowest resistance was related to piperacillin (60.6%) and carbapenems (14.6%) respectively. 80.5% (33 of 41) isolates were positive by MHT, but all of them (100%) were negative for amplification of the bla-KPC gene in the PCR method.
    Conclusion
    The MHT was an appropriate method for approving carbapenemase production. Moreover, a laboratory could accept the carbapenemase production with PCR method for the bla-KPC gene, which has the additional profit of validating which KPC is present.
    Keywords: Klebsiella pneumoniae, Carbapenem Resistance, bla, KPC gene, MHT
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