PCR vs. Conventional Methods: Diagnosing Trichomonas vaginalis Infections | InformativeBD

Dr. Kiranmai, and Dr. A. Neelima, from the different institute of the india. wrote a Research Article about, PCR vs. Conventional Methods: Diagnosing Trichomonas vaginalis Infections. Entitled, Evaluation of pcr in the molecular diagnosis of trichomonas vaginalis infection in comparison with other conventional methods. This research paper published by the International journal of Microbiology and Mycology (IJMM). an open access scholarly research journal on Microbiology. under the affiliation of the International Network For Natural Sciences| INNSpub. an open access multidisciplinary research journal publisher.

Abstract

Trichomonas vaginalis (T. vaginalis) is a common pathogen with worldwide distribution. It is estimated that worldwide 180 million people are infected annually. Trichomoniasis is associated with vaginitis, cervicitis, low birth weight, and preterm delivery. PCR has the advantage of high sensitivity, shorter time for diagnosis and the ability to detect nonviable or defective organism. In this study we used these three methods for evaluation of PCR in comparison with conventional methods like wet mount and culture in the detection of T. vaginalis in vaginal discharge. Three vaginal swab specimens were obtained from each of 200 cases, of the age group 18-40years, both symptomatic and asymptomatic females attending Gynaecology OPD(50) and Family planning OPD(50) at Gandhi hospital, Secunderabadand two FSW(Female sex workers) clinics (100) in highly concentrated areas of them in Hyderabad, for validation of various forms of Trichomonas vaginalis diagnostic procedures. One swab was immediately examined by wetmount microscopy, a second swab was placed in Wittington’s medium for cultivation, and other swab is placed in 2SP transport medium for PCR for T.vaginalis. A total of 58 samples positive in one or more tests were identified: 11 (5.5%) infections were detected by wet mount microscopy, and 30 (15%) positives in culture respectively. PCR was positive in 50 (25%) samples. PCR appears to be the most sensitive method with high detection rate and method of choice for detection of genital infections with T. vaginalis.

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Introduction

Worldwide, Trichomonas vaginalis causes approximately 180 million new infections per year, making it the most prevalent non-viral sexually transmitted disease (STD) agent. (Petrin et al., 1998; Kengue et al., 1994; Madico et al., 1998).

Infections in women can cause vaginitis, urethritis, and cervicitis, and complications include premature labor, lowbirth-weight offspring, and post abortion or post hysterectomy infection (Shaio, et al., 1997). It has been estimated that 10 to 50% of T. vaginalis infections in women are asymptomatic and in men the proportion may even be higher. (Burstein, G. R et al., 1999) This parasite has also been implicated as a cofactor in the transmission of the human immunodeficiency virus and other nonulcerative STD agents. However, since the incidence of T. vaginalis infection is highest for groups with a high prevalence of other STDs, this latter hypothesis remains to be confirmed (Madico et al., 1998). In addition, a relationship between T. vaginalis infec-tion and cervical cancer has recently been suggested (Zhang et al., 1995). 

The most common tool for diagnosis of T. vaginalis infection is still microscopic examination of wet mount preparations, which has a sensitivity of approximately 38-60%. (Fouts et al., 1980) Microscopic examination of cultures of the parasite in specialized mediaimproves the sensitivity to 70-85% (Gelbart et al., 1990; Schmid et al., 1989). However, the quality of these diagnostic tests is strongly dependent on theskills and experience of the microscopist and also on the quality of the sample. Diagnosis traditionally depends on the microscopic observation of motile protozoa in vaginal discharge. Culture requires a special medium and the result takes up to 7 days. Diagnostic improvements have been suggested since years. Finally, molecular techniques such as fluorescent in situ hybridization, oligonucleotide probing, and PCR have been developed. (Petrin et al., 1998; Kengue et al., 1994; Zhang et al., 1995; Muresu et al., 1994; Philips Heine et al 1997; Riley et al., 1992; Rubino et al., 1991).

To date, numerous T. vaginalis specific PCR assays have been described. Examples of targets include the ferridoxin gene, beta tubulin gene, highly repeated DNA sequencing and 18s ribosomal genes. (Riley et al., 1992; Kengue et al., 1994; Madico et al., 1998; Mayta et al., 2000). 

Marcia M. Hobbs et al., compared culture and PCR ELISA in urethral swabs, urine and semen for TV detection in male sexual partners of women with trichomoniasis identified by wet mount and culture. TV was detected more often in men with wet mount positive partners emphasizing the importance of partner evaluation and treatment. Even with a sensitive PCR assay, reliable detection of TV in male partners required multiple specimens.

Charlotte Gaydos et al., (2006), compared Gene Probe transcription mediated amplification TV research assay and real time PCR for TV detection using a Roche Light Cycler instrument with female self-obtained vaginal swab samples and male urine samples. The Gen Probe TMA assay is commercially available as an analyte specific reagent and offers laboratories a highly sensitive and specific assay for use clinically.

Rasoul Jamali et al., (2006), compared diagnosis of Trichomonas vaginalis infection using PCR method to culture and wet mount microscopy and concluded that PCR had high sensitivity and slightly less specificity than wet mount taking culture as the standard. 

This study was done to diagnose both symptomatic and asymptomatic female patients with Trichomonas vaginalis infection by Microscopy, Culture and PCR and to compare the efficacy of each of the above diagnostic modalities in terms of rapidity, sensitivity and specificity.

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