Pr. Lin et al., ONE-TUBE, NESTED-PCR ASSAY FOR THE DETECTION OF TRICHOMONAS-VAGINALISIN VAGINAL DISCHARGES, Annals of tropical medicine and parasitology, 91(1), 1997, pp. 61-65
To shorten the time and improve the accuracy of diagnosis of vaginal t
richomoniasis, a novel, one-tube, nested PCR, which targets a family o
f 650-bp specific DNA repeats from the Trichomonas vaginalis genome, h
as been developed. Samples were prepared by a rapid boiling method and
the PCR products analysed by gel electrophoresis. A 290-bp DNA fragme
nt was observed in all positive cases. No cross-reaction with any othe
r pathogens, including the Pentatrichomonas hominis and Giardia lambli
a used as controls, was found. Using the assay, one genome-equivalent
of T. vaginalis in 20 mu l vaginal discharge can be detected and diagn
osis can be made within 6 h. When 165 clinical specimens were examined
by wet mount, culture and the PCR assay, 16 were found positive for T
. vaginalis by both culture and PCR, whereas only nine of these 16 cas
es were found to be positive by examination of met mounts. No PCR-nega
tive cases were positive by wet mount or culture. This new assay appea
rs to be a simple, rapid, accurate and sensitive method for the diagno
sis of vaginal trichomoniasis.