Abstract
BackgroundTrichomoniasis resulting from Trichomonas vaginalis (T. vaginalis) has been considered as a commonly seen disease with the transmission way of sex. At present, the detection methods of T. vaginalis mainly include wet mount microscopy, culture, PCR, immunofluorescence and ELISA. However, all of these detection methods exist shortcomings.MethodsIn this study, a loop-mediated isothermal amplification (LAMP) assay that targeted the species-specific sequence of adhesion protein 65 (AP65) gene had been conducted to detect T. vaginalis. The optimum reaction system and conditions were optimized in this rapid detection method.ResultsThe results of sensitivity analysis showed that the LAMP assay targeting the AP65 gene was 1000 times more sensitive than the nested PCR targeting the actin gene commonly used for detection of T. vaginalis, and the detecting limitation of the former was 10 trichomonad. Moreover, the amplification of the target gene AP65 by LAMP assay exhibited high specificity and the product was exclusively from T. vaginalis. The detection technique of LAMP did not exhibit cross-reactivity with the common pathogens of Trichinella spiralis, Toxoplasma gondii, Escherichia coli, Candida albicans, Staphylococcus aureus, Haemophilus.ConclusionsAccording to the present study, the LAMP assay with the target of AP65 gene, was suitable for the early diagnosis of T. vaginalis infections. Consequently, the LAMP assay was proposed by the current study as a point-of-care examination and an alternative molecular tool which exhibited the potential value in the treatment, control and prevention of trichomoniasis transmission and relevant complication.
Highlights
Trichomoniasis resulting from Trichomonas vaginalis (T. vaginalis) has been considered as a commonly seen disease with the transmission way of sex
We developed a novel way to detect T. vaginalis based on loop-mediated isothermal amplification targeting adhesion protein 65
Detecting T. vaginalis by nested Polymerase Chain Reaction (PCR) and loop-mediated isothermal amplification (LAMP) T. vaginalis actin gene was amplified using nest PCR with specific primers, and a positive sequence of about 1100 bp was obtained in line 3 and 4 with the expectation (Fig. 1a)
Summary
Trichomoniasis resulting from Trichomonas vaginalis (T. vaginalis) has been considered as a commonly seen disease with the transmission way of sex. The detection methods of T. vaginalis mainly include wet mount microscopy, culture, PCR, immunofluorescence and ELISA. All of these detection methods exist shortcomings. More than 170 million people were infected with the disease every year in the world, and the infection rate of people around the world was different and had an increasing trend [9]. In the United States, nearly 5 million people were infected with T. vaginalis every year, while the infection rate was 24.3% in Japan, 23.8% in Uganda and 18.0% in South Africa [10]. The widespread infection and prevalence of T. vaginalis increased the risk of human infection with human immunodeficiency virus (HIV) and mycoplasma, and T. vaginalis has become an important co-factor of HIV infection [20]
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