International Journal of Molecular and Clinical Microbiology
Volume:2 Issue: 2, Summer and Autumn 2012

Plasmids play an important role in the virulence and antibiotic resistance of Salmonella. Salmonella carry plasmids having different size and number. Plasmid profile analysis of single isolates of Salmonella typhimurium and Salmonella enteritidis revealed the presence of 4- plasmid profiles. Salmonella serovars produce several Type III secretions including Sop-E and Sop –B. Soap E is detected by Dot-ELISA. Dot-ELISA is one of the solid phase immuneassay used to detect either antigens or antibody. Sop-B (Salmonella outer protein-B) is detected by multiplex PCR method using PCR primers Sop P1 and SOP- M13. It is concluded that the technique used in this study is more sensitive and identification and determination of various molecular characteristics of two species enhances the knowledge towards the vaccine production.

Candida albicans is an opportunistic fungi that is able to thrive in many host niches, including the skin, mucosal, surfaces, the blood stream and internal organs. Agglutinin-like sequence (ALS) genes which could play a role in forming biofilms, adherence to host surfaces as a virulence factor and antifungal drug resistant. The purpose of the present study was to evaluate the presence ALS1 and ALS3 genes in clinical strains of C.albicans isolated from women with vaginal candidiasis symptom. Vaginal swabs were collected from patients suffering from vaginal candidiasis and admitted to health care centers (Tehran, Iran) from June 2011 until June 2012 and cultured on Sabaouraud Dextros Agar for 48 hours. A PCR-RFLP was used with MspI restriction enzyme for identification of Candida albicans. Susceptibility testing of 53 clinical isolates of C.albicans isolates was done against Fluconazole by using disk diffusion method. Total DNA was extracted from C.albicans isolates and PCR assay was used to evaluate the presence of ALS1, ALS3 and internal control (ACT1) genes. 53 clinical isolates out of 100 were identified as C.albicans by using PCR-RFLP. Three (5.7%) of them were susceptible and 50 (94.3%) were resistant to fluconazole. 44 patients (83%) with vaginal candidiasis were positive for ALS1 gene and 48 ones (90.5%) were positive for ALS3. According our finding, a significant correlation was seen between the presence of ALS1 and ALS3 genes and fluconazole resistance in C.albicans isolates.

Cytomegalovirus (CMV) is a widespread pathogen that is found in milk, saliva, urine, cervical secretions, and semen fluid. It is supposed that CMV is a risk factor for breast cancer. The aim of this study was to determine the frequency of CMV among benign and malignant breast tumors. Paraffin embedded breast carcinoma (n=24) and fibroadenoma (n=24) samples were collected from Toos and Firoozgar hospitals in Tehran, Iran, during the year 2012. PCR technique was used to detect CMV genome in the samples following the DNA extraction. Out of 24 carcinoma samples, 2 (8.3%) were positive for CMV, while no CMV positive sample was found among fibroadenoma tumors. Statistic analysis showed no significant correlation between this virus and formation of either benign or malignant tumors of breast. The role of CMV in a number of human cancers has been shown during the past decade. This study shows no relationship between CMV infection and breast benign and malignant tumors. However, future studies are needed to better understand the role of this virus in formation of breast tumors.

In the present study, variation among different bacterial strains was observed in the plaques of tobacco chewers (TC) as compared to those from normal persons (non tobacco chewers) (NTC). Bacterial strain J1 (dominant in the plaques of NTC) did not initiate the process of adhesion (biofilm formation) until it appeared to have reached a required population density necessary for the production of adhesive substances by the bacterium, which indicating the involvement of quorum-sensing mechanism. The spent media of bacterial strain J15 (dominant in the plaques of TC) also substantially reduced the adhesion of the respective bacteria to the acrylic particles by bacterial strain J1. This indicates the possibilities of existence of some quorum-quenching molecules in the spent medium of strain J1 as well as strain J15 bacteria. These molecules might be exploited for efficient management of dental plaque-biofilms. It may help to avoid the use of extremely high doses of antibiotics which may be harmful in the long run.

Today, chronic viral hepatitis is considered as a public health problem in the world. Totally, 5% of the world population, especially in Asia and Africa suffer from this problem. The prevalence of acute hepatitis in America is 0.1-0.2 percent, 50% of those with hepatitis C (HCV) and 5-10% of them with hepatitis B (HCV) develop chronic hepatitis. Determining HBV and HCV genotypes is important to clarify the disease and pathogenicity of the virus since genotypes are different in terms of pathogenicity and response to treatment. In this project, the number of 90 patients with hepatitis B and C referred to a private medical diagnostic laboratory in Rasht was included in this study. After obtaining blood samples from patients, genotyping and determining the amount of hepatitis B and C virus were performed. It was found that out of 90 patients, 56 ones (62.2%) had HBV and 34 (37.8%) HCV. In all 56 patients with genotype HBV and in 34 patients with genotype HCV observed genotypes including 1a, 1b, and 3a. The most frequent genotype was 1a which was seen in 18 patients (20%) followed by 3a in 14 patients (15.6%). Only 2 patients (2.2%) had genotype 1b. Knowing the genotype and the viral load of HBV and HCV in each region is important in disease development and treatment. Determining HBV and HCV genotypes is important to clarify the disease progression and pathogenicity of the virus.

The rate of cervical cancer in Asians is deferent, based on the risk factors affecting and the rate of papiloma virus infection among females. Cervical cancer is one of the important cancers between Iranian women. In the present study the rate of cervical neoplasia was detected in Isfahan and some risk factors affecting the incidence of cervical cancer was analyzed. The human papiloma virus infection was also detected in cervical neoplasia via PCR method after DNA extraction. 15000 pap smear tests results achieved from patient documents for detection of the rate of squamous cell neoplasia, based on at least one report in pap smear test in a random and distributed sampling. We have also studied the effect of some risk factors on the incidence of cervical cancer in 100 patient and 100 females without any neoplasia report as a control group. The results analyzed by Chi2 and T tests. The rate of human papiloma virus (HPV) infection detected in pap smear specimens using specific primers for amplification of HPV genome. At least one neoplastic report was shown in 0.91% of females and 0.61% of which had at least one ascus report. 55.4% of patients with neoplasic pap tests were infected with HPV in PCR test. Teenage marriage, high parities, lower education and smoking increases the risk of cervical neoplasia. It is concluded that molecular methods including genome analysis are useful method in identification of HPV.

Salmonella enterica serotypes are one of the most important food borne pathogens and significant public health concerns around the world in humans and other animal species. A total of eighty three epidemiologically unrelated clinical isolates of Salmonella enterica serovars were subjected to antimicrobial susceptibility testing. Eleven isolates (13.1%) which were resistant to at least 4 groups of antimicrobial agents considered as multidrug resistant (MDR) Salmonella serovars. Emergence of MDR Salmonella serovars demonstrates that antimicrobial selection pressure is widespread in our clinical settings. According to the results of antimicrobial susceptibility testing, Salmonella clinical isolates are more susceptible to fluoroquinolones and third generation cephalosporins and these drugs may be used as drugs of choice to treat Salmonella infections.

H.pylori is one of the common infectious agent that involves in acute and chronic gastritis, peptic scar and gastric cancer. The infection episode in children ranges between 10 to 80 percent and the high frequency in developing countries. The goal of this study was to determine the incident of H.pylori in the patients carried out to endoscopy and the effective factors. In this study, 50 patients of 3 to 15 years old range that were admitted in endoscopy ward were studied. After endoscopy, the specimen of gastric mucus was studied using rapid urease Test (RUT). The specimen with urease positive test were taken as the case and specimen with negative urease test were taken as control group and the factors involves in H.pylori were studied. The Odds ratio was calculated in the specimen and confidence interval was performed in 95% probability and the data was analyzed by using common statistical tests. The results showed that of 50 children, 9 patients (18%) were diagnosed with Pelvic ulcer disease (PUD) and positive H.pylori, 5 patients (10%) had gastritis and free from contamination to concerned organism. The 6-15 years old age group showed the highest number of bacteria (30%). Of all the patients subject of study, 7 cases were male (14%) and 8 (16%) were females. 25 patients (54%) had less than high school diploma education and 23 cases (46%) had university education. In this study, age, sex, water consumption, number of family members and the economic-social status were the criteria of study and it was shown that the infection essentially happens in young ages and the chance of infection through contaminated water is high. It is concluded that H.pylori infection is an important factor involving the gastritis and duodenum inflammation.

Candida albicans is the most common and important factor causing candidiasis infections in human. As fungi are resistant to anti-fungal agents, and also with respect to the restrictions in treatment of fungal diseases such as their high costs and side effects, study of the combination of herbal medicine is truly required. The aim of the present study was to investigate the effects of herbal essences of eucalyptus and lavandula on the growth of clinical Candida albicans strains which have been separated from mucosal, cutaneous and visceral infections, and compared to those Candida albicans which are resistant and sensitive to caspofungin. The present study has been carried out on 41 Candida albicans taken from patients suffering from mucosal, cutaneous and visceral candidiasis. The degree of samples’ sensitivity to eucalyptus and lavandula were determined through applying broth microdilution method. In the present study, the MIC (minimum inhibitory concentration) of eucalyptus against Candida albicans was determined at 4-1024µl/ml, in which the most growth fluctuations were seen in the densities of 512 and 128 µl/ml. Moreover, the MIC of lavandula was determined at 1-256 µl/ml, and the most fluctuations in its growth were in the density of 32 and 16 µl/ml. Eucalyptus and lavandula essences have a very good anti-fungal effect against Candida albicans. In the present study, lavandula essence proved better anti-candidiasis effects while no considerable difference was seen between the strains sensitive and resistant to caspofungin in its MIC. In higher densities, eucalyptus essence prevents strains from growing. Therefore, it is recommended to investigate the effects of these essences on other pathogenic fungi and bacteria. It is also recommended to evaluate the use of combinations of two or more essences for curing infections or preventing their reoccurrence.

The lack of variation in antifungal drugs, and the misuse or inappropriate use usually causes resistant strains of the yeast in human’s normal flora. Unfortunately, a large number of Candida infection cases in immunosuppressed patients with insufficient treatment eventually can cause patient's death. The aim of this study was to evaluate the in vitro conventional antifungal azole compounds with binary mixture with appropriate ratios by susceptibility test using the laboratory as a mixture of two in vitro conditions. In present study, 10 isolates of Candida were admitted from patients that were referred to mycology laboratory of Faghihi Hospital at Shiraz with signs of cutaneous and mucosal infections. In the present study we used the methods of the binary mixure of common antifungal drugs Clotrimazole, Ketoconazole and Miconazole on equally proportion have been used. The drugs were solved in various concentrations on the SDA medium and then Cndida isolates were cultured in the SDA plates. The minimum inhibitory concentration (MIC 90) and minimum fungicidal concentration (MFC) was determined. The results of this study suggest that a binary mixture of these drugs inhibited the growth of most strains of pathogenic Cndida isolates. The combination with Clotrimazole and ketoconazole in equal proportion had more effective than other drug mixtures against all isolates with the exception of isolate 3. In contrary, the combination of Miconazole and Clotrimazole had the least effect, and the MIC was calculated in the range of 3.12 to 50 μg/ml. Evaluation of MFC showed almost the same results. Lowest values of the MFC belonged to the combination of clotrimazole and ketoconazole which was obtained 6.25 μg/ml. It is concluded that the use of the combination of Clotrimazole and Ketoconazole in equal ratios has better antifungal effects against cutaneous and mucosal isolates of the Candida infection.