Evaluation of a Polymerase Chain Reaction (PCR) Assay for the diagnosis of Trichomonas Vaginalis Infection.

Hema, Paul (2009) Evaluation of a Polymerase Chain Reaction (PCR) Assay for the diagnosis of Trichomonas Vaginalis Infection. Masters thesis, Christian Medical College, Vellore.


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INTRODUCTION : Trichomonas vaginalis is a protozoan parasite and etiologic agent for Trichomoniasis, a sexually transmitted disease (STD). Trichomoniasis is a common non-viral sexually transmitted disease. The global incidence of trichomoniasis is 174 million annually. In South and South East Asia, 76.5 million new cases are reported annually. Symptomatic trichomoniasis is more common in women than in men. Trichomoniasis is most prevalent during the peak years of sexual activity. The route of transmission is almost exclusively by sexual intercourse. It has been estimated that 10-50% of Trichomonas vaginalis infection in women is asymptomatic. Trichomoniasis has a wide spectrum of symptoms ranging from inflammation in the form of vaginitis, urethritis to malodourous frothy discharge alone with asymptomatic carrier state. The discharge is frothy yellow or green and mucopurulent. About 2% of infected patients present with "strawberry cervix". Trichomoniasis is associated with serious complications. These include pyosalpinx, endometritis, and premature rupture of membrane, preterm labor and low birth weight. Trichomonas vaginalis is associated with non gonococcal urethritis and prostatitis in men. AIM AND OBJECTIVES : AIM : To standardize and evaluate a Polymerase Chain Reaction (PCR) for the diagnosis of Trichomoniasis. OBJECTIVES : 1. To standardize Polymerase Chain Reaction (PCR) for the diagnosis of Trichomonas Vaginalis in culture from clinical samples. 2. To evaluate the performance of the PCR in diagnosing trichomoniasis in the hospital Setting. 3. To determine the prevalence of trichomoniasis in Human Immunodefiency Virus (HIV) Positive women. MATERIAL AND METHODS : The study protocol was approved by the institutional review board. The study had two parts. The first part of the study was to standardize a non-nested PCR for the amplification of the one of the Trichomonas vaginalis gene and to evaluate its performance on frozen culture positive and culture negative samples. The samples for these culture supernatants were collected from women, who had presented with vaginal discharge. These women belonged to a rural area looked after by the community department of Christian Medical College. The vaginal swabs from these women were collected between the year 2005- 2006 and inoculated into modified Diamond’s media. RESULTS : The first set of primers (TVA5/ TVA6) amplified the Trichomonas vaginalis DNA from all the three samples i.e., undiluted supernatant, undiluted concentrate and 1/10 diluted concentrate. The expected band size for the product was 102bp. The second set of primers, Beta tub9/2 also showed amplification with all the three samples with different dilutions used. However, the undiluted concentrate showed a very weak band. PCR was repeated with same sample and primer set Beta tub9/2 where two concentrations of primers were tried, 25mM/ reaction and 10mM/ reaction, the concentration of other reagents as well as cycling conditions remained same. There was no mprovement in the results. The gel picture showing the Trichomonas vaginalis specific bands with different concentrations of samples with the two sets of primers used. CONCLUSION : Trichomoniasis caused by the protozoan parasite Trichomonas vaginalis is the most common non-viral sexually transmitted infection. In this study reported here a PCR targeting the adhesion and Beta tubulin genes of T. vaginalis was standardized for the detection of the parasite in vaginal swab samples. The usability of this PCR was compared with culture and wet preparation of prospectively collected vaginal swab samples from HIV infected women attending infectious disease clinics.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Polymerase Chain Reaction ; Evaluation ; Assay ; Diagnosis ; Trichomonas Vaginalis Infection.
Subjects: MEDICAL > Microbiology
Depositing User: Subramani R
Date Deposited: 14 Aug 2017 02:00
Last Modified: 14 Aug 2017 02:00
URI: http://repository-tnmgrmu.ac.in/id/eprint/1332

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