Trichomonas vaginalis: repeated DNA target for highly sensitive and specific polymerase chain reaction diagnosis.

Important note: Information in this article was accurate in 1995. The state of the art may have changed since the publication date.

Trichomonas vaginalis: repeated DNA target for highly sensitive and specific polymerase chain reaction diagnosis.

Cell Mol Biol (Noisy-le-grand). 1994 Sep;40(6):819-31. Unique Identifier : AIDSLINE GENBANK/L23861
Kengne P; Veas F; Vidal N; Rey JL; Cuny G; Laboratoire Retrovirus-Parasites, Institut Francais de; Recherche Scientifique pour le Developpement en Cooperation; (ORSTOM), Montpellier, France.

Abstract: Trichomoniasis is recognised as a major sexually transmitted disease (STD) in the world and may act as an acquired immunodeficiency syndromes (AIDS) co-factor by enhancing the transmission of human immunodeficiency virus (HIV). Diagnosis of Trichomonas vaginalis can be achieved by several methods, but sensitive detection means are still lacking. In this study a 2000-bp repeated DNA fragment of T. vaginalis was cloned. Part of a conserved region of this insert was sequenced, two primers (TVK3 and TVK4) were chosen and a highly sensitive detection by polymerase chain reaction (PCR) was then developed for T. vaginalis. All strains of T. vaginalis analysed with these primers gave the expected 350-bp fragment and a 450-bp additional fragment. Sequence analysis of these PCR amplification products revealed that the 450-bp fragment contained the 350-bp with a 100-bp insertion characterised by a TGG microsatellite. A second primer set, namely TVK3 and TVK7 (determined at the border of the insertion), yielded PCR products of expected sizes. After amplification we were able to detect a single parasite. We also detected T. vaginalis in vaginal fluids of patients with STD. There was no reaction with human DNA or other infectious agents. It appears that the two set primers are highly specific of T. vaginalis and provide a useful tool for PCR diagnosis in asymptomatic and symptomatic patients especially among the HIV at risk individuals.

Keywords: Animal Base Sequence Blotting, Southern Consensus Sequence DNA Insertion Elements DNA Primers DNA, Protozoan/CHEMISTRY/*ISOLATION & PURIF DNA, Satellite/CHEMISTRY/*ISOLATION & PURIF Female Human Molecular Sequence Data Polymerase Chain Reaction/*METHODS *Repetitive Sequences, Nucleic Acid Sensitivity and Specificity Trichomonas vaginalis/GENETICS/*ISOLATION & PURIF Trichomonas Vaginitis/*DIAGNOSIS JOURNAL ARTICLE
950430
M9540710

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