A study of microsporidiosis in corneal scrapings of keratitis patients referring to Farabi Eye Hospital, Tehran, Iran in 2013-14.

BACKGROUND AND PURPOSE: Microsporidiosis is one of the emerging and opportunistic infections, which causing various clinical symptoms in humans. The prevalence of this infection varies, depending on the infected organ, diagnostic methods, and geographical conditions. In the present study, we aimed to investigate microsporidial keratitis in patients referring to Farabi Eye Hospital Tehran, Iran in 2013-14. MATERIALS AND METHODS: Two scraping samples were collected from 91 keratitis patients, five cases had prior history of receiving immune suppressive drugs. One of the two collected samples from each participant was used for Vero cell culture and the other was used for the preparation of Giemsa and Gram staining slides. After 30 days, the cells were scrapped and used for DNA extraction; afterwards, nested polymerase chain reaction (PCR) detection method was applied. Primer pairs of small-subunit ribosomal RNA gene were designed by CLC Genomics workbench software to amplify all major microsporidian pathogens, as well as E. bieneusi , which was used as the positive control in this study. RESULTS: The nested PCR showed negative results regarding the presence of microsporidia in the samples. Similarly, Giemsa and Gram staining slides did not detect any spores. CONCLUSION: The prevalence of human microsporidiosis ranges between 0% and 50%, worldwide. Based on all the negative samples in the present study, we can conclude that the prevalence of this infection among Iranian patients falls in the lower quartile. By gathering further evidence, researchers can take a step forward in this area and open new doors for the assessment of AIDS patients and users of immunosuppressive drugs.

Double-Stranded RNA Viral Infection in Tehran Trichomonas vaginalis Isolates.

BACKGROUND: Trichomonas vaginalis is a pathogenic protozoon and may be contaminated with dsRNA virus called Trichomonas vaginalis virus (TVV). The viral infection is an important factor for its pathogenesis and sensitivity to metronidazole. The presence of TVV is associated with qualitative and quantitative expression of cysteine proteinases and surface immunogenic; P270. The purpose of this study was to determine TVV frequency in T. vaginalis clinical isolates in Tehran, Iran. METHODS: The 46 T. vaginalis isolates were collected from Tehran Province and cultured in TYI-S-33 culture medium. Viral RNA was extracted and RT-PCR was done. RESULTS: Of 46 T. vaginalis isolates, 8 isolates (17.39%) were infected with TVV-1. There was not any association between patient age and TVV- infected T. vaginalis. There were 17.39% viral infection in T. vaginalis isolates which was lower than that reported by other researchers. CONCLUSION: This is the first report on T. vaginalis isolates infection by TVV-1 in Iran.

Gene Diversity of Trichomonas vaginalis Isolates.

BACKGROUND: Trichomonas vaginalis is protozoan parasite responsible for trichomoniasis and is more common in high-risk behavior group such as prostitute individuals. Interest in trichomoniasis is due to increase one's susceptibility to viruses such as herpes, human papillomavirus and HIV. The aim of this study was to find genotypic differences between the isolates. METHODS: Forty isolates from prisoners' women in Tehran province were used in this study. The random amplified polymorphic DNA (RAPD) technique was used to determine genetic differences among isolates and was correlated with patient's records. By each primer the banding pattern size of each isolates was scored (bp), genetic differences were studied, and the genealogical tree was constructed by using NTSYS software program and UPGMA method. RESULTS: The least number of bands were seen by using primer OPD8 and the most by using OPD3. Results showed no significant difference in isolates from different geographical areas in Iran. By using primer OPD1 specific amplified fragment with length 1300 base pair were found in only 8 isolates. All these isolates were belonged to addicted women; however, six belonged to asymptomatic patients and two to symptomatic ones. CONCLUSION: There was not much genetic diversity in T vaginalis isolates from three different geographical areas.

Detection of Neisseria gonorrhoeae from vaginal swabs of Ewin, Rajaii shahr, Karaj and Varamin female prisoners by PCR and culture methods.

Isolation of N. gonorrhoeae by culture method is currently the gold standard for the definitive diagnosis of gonorrhoea. However, PCR techniques are being used more frequently as sensitivity and specificity of the newer tests are improved. In this study, 500 vaginal swabs from Ewin, Rajaii shahr, Karaj and Varamin female prisoners were used for detection of N. gonorrhoae by culture and PCR techniques. Five hundred vaginal swabs from Ewin, Rajaii shahr, Karaj and Varamin female prisoners were cultured in modified Thayer Martin in 37 degrees C with 5% CO2 for 72 h. Oxidase, catalase tests, biochemical tests such as maltose and glucose oxidation and gram staining, were used to confirm the isolated species. Amplification by PCR using 2 targets which are specific for N. gonorrhoeae, Ngu1 and Ngu2, were used to detect the presence of gonococcal specific DNA. Despite of finding some questionable samples as N. gonorrhoeae by using biochemical tests, PCR method confirmed that none of them were positive for N. gonorrhoeae. This study deals with detection of N. gonorrhoeae among woman prisoners in three main prisons in Tehran, Iran. The high specificity and sensitivity coupled with low cost and rapidity of the method (PCR) provided a substantial advantages over the time consuming culture methods currently used in hospitals and laboratories.

Molecular diagnosis of trichomoniasis in negative samples examined by direct smear and culture.

BACKGROUND: Trichomoniasis is an extremely common sexually transmitted infection (STI) worldwide and is associated with important public health problems, including amplification of HIV transmission. This disease is in forms of symptomatic and asymptomatic in women and may depend on host as well as parasite variables. Most of the studies reported from females are based on examination of vaginal secretions and urine samples by direct smear and culture in modified Diamond's media. The aim of this study was checking the samples, which were negative by direct smear and culture, with PCR technique. METHODS: The urine samples and vaginal discharge of patients attending Gynecology Clinics of Mazandaran Province, Iran with different symptoms rechecked for Trichomonas vaginalis by PCR technique using primers targeting a conserved region of the beta-tubulin genes of the parasite. Data were analyzed by Epi Info software program RESULTS: Out of 161 negative samples by direct smear and culture, seven samples (4.3%) were positive by PCR technique. CONCLUSION: Diagnosis of trichomoniasis by PCR is a sensitive and specific method that could play important role to help the physicians for properly treatment and control of infection.