Loop mediated isothermal amplification assay for detection of Trichomonas vaginalis in vaginal swabs among symptomatic women from North India

Abstract

Significance and Impact of the Study: Trichomonas vaginalis is a common sexually transmitted pathogen associated with considerable morbidity and risk of complications. Due to the limitations of traditional diagnostic modalities, three molecular assays were compared: conventional polymerase chain reaction (PCR), Xpert TV assay, and loop mediated isothermal amplification (LAMP) assay for detecting T. vaginalis in symptomatic females. All tests had a sensitivity of 100% and the inter-rater reliability was excellent AbstractTrichomonas vaginalis is one of the most common curable sexually transmitted pathogens infecting both men and women worldwide. Unlike traditional methods such as microscopy and culture, nucleic acid amplification tests rapidly detect this agent, assisting in treatment. Conventional polymerase chain reaction (PCR), the loop-mediated isothermal amplification (LAMP), and the Xpert TV assay were evaluated using 28 microscopy positive T. vaginalis samples and 125 microscopy negative samples from symptomatic females of reproductive age. The sensitivity of all tests was 100% and the specificity was 100%, 100%, and 99Á2% for PCR, Xpert TV, and LAMP, respectively. The inter-rater reliability was excellent for PCR: Xpert TV (kappa-coefficient = 1) and good for LAMP assay: Xpert TV/PCR (kappa-coefficient = 0Á98) and conventional PCR: LAMP (kappa-coefficient = 0Á98). The study highlights the importance of PCR for screening T. vaginalis in women, particularly in laboratories where the Xpert-TV assay is not available or not affordable. The LAMP assay showed a lower positive predictive value which merits further evaluation.

Prevalence of nonviral reproductive tract infections/sexually transmitted infections in female patients with cervicovaginal discharge

“…The second swab (cervical or cervicovaginal) was used to inoculate onto chocolate agar and it was then squeezed thoroughly into a falcon tube containing molecular grade water for DNA extraction. The DNA was extracted using Chelex® (Sigma Aldrich, MO, USA) and amplified for detecting the presence of TV, Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) using polymerase chain reactions (PCRs), as described earlier[ 10 11 ] [ Supplementary File ]. The chocolate agar and blood agar plates were incubated in a CO 2 incubator (Binder, GmbH Germany) at 37°C for at least 72 h, the plates were examined for the presence of any visible growth.…”

“…and Ureaplasma urealyticum . The urine specimens obtained were subjected only to PCR, as described earlier[ 10 11 ] [ Supplementary File ].…”

A retrospective analysis of sexually transmitted infections among males presenting to a tertiary care hospital of India

“…The second swab was used to put up NG and CT PCR using three conventional PCRs, as described earlier. [ 5 6 ] The swab was thoroughly squeezed in molecular grade water for DNA extraction. DNA was extracted using Chelex® (Sigma Aldrich, MO, USA) and amplified.…”

A microfluidic-chip-based system with loop-mediated isothermal amplification for rapid and parallel detection of Trichomonas vaginalis and human papillomavirus

“…And as far as we know, our work has been the first to simultaneously detect T. vaginalis and different types of HPV with LAMP and microfluidic-chip technology, though several studies have reported detection of T. vaginalis or HPV with LAMP or combined with microfluidic chips. 18,34 There have been many studies employing microfluidic systems with LAMP for nucleic acid detection. 19,35,36 We compared our microfluidic-chip-based system with previous reported works, in particular to those targeting related pathogens using LAMP (Table S3 †).…”