Loop-Mediated Isothermal Amplification Targeting Actin DNA of Trichomonas vaginalis
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초록
Trichomoniasis caused by Trichomonas vaginalis is a common sexually transmitted disease. Its association with several health problems, including preterm birth, pelvic inflammatory disease, cervical cancer, and transmission of human immunodeficiency virus, emphasizes the importance of improved access to early and accurate detection of T. vaginalis. In this study, a rapid and efficient loop-mediated isothermal amplification-based method for the detection of T. vaginalis was developed and validated, using vaginal swab specimens from subjects suspected to have trichomoniasis. The LAMP assay targeting the actin gene was highly sensitive with detection limits of 1 trichomonad and 1 pg of T. vaginalis DNA per reaction, and specifically amplified the target gene only from T. vaginalis. Validation of this assay showed that it had the highest sensitivity and better agreement with PCR (used as the gold standard) compared to microscopy and multiplex PCR. This study showed that the LAMP assay, targeting the actin gene, could be used to diagnose early infections of T. vaginalis. Thus, we have provided an alternative molecular diagnostic tool and a point-of-care test that may help to prevent trichomoniasis transmission and associated complications. Key words: Trichomonas vaginalis, trichomoniasis, PCR, multiplex PCR, loop-mediated isothermal amplification
Abstract
Trichomoniasis caused by Trichomonas vaginalis is a common sexually transmitted disease. Its association with several health problems, including preterm birth, pelvic inflammatory disease, cervical cancer, and transmission of human immunodeficiency virus, emphasizes the importance of improved access to early and accurate detection of T. vaginalis. In this study, a rapid and efficient loop-mediated isothermal amplification-based method for the detection of T. vaginalis was developed and validated, using vaginal swab specimens from subjects suspected to have trichomoniasis. The LAMP assay targeting the actin gene was highly sensitive with detection limits of 1 trichomonad and 1 pg of T. vaginalis DNA per reaction, and specifically amplified the target gene only from T. vaginalis. Validation of this assay showed that it had the highest sensitivity and better agreement with PCR (used as the gold standard) compared to microscopy and multiplex PCR. This study showed that the LAMP assay, targeting the actin gene, could be used to diagnose early infections of T. vaginalis. Thus, we have provided an alternative molecular diagnostic tool and a point-of-care test that may help to prevent trichomoniasis transmission and associated complications. Key words: Trichomonas vaginalis, trichomoniasis, PCR, multiplex PCR, loop-mediated isothermal amplification
참고문헌 (33)
- Gerbase A, "Global prevalence and incidence estimates of selected curable STDs", Sex Transm Infect, 74(suppl), pp.S12~S16 (1998)
- Rowley J, "Global incidence and prevalence of selected curable sexually transmitted infections-2008" (2012), WHO
- Fichorova RN, "Impact of T. vaginalis infection on innate immune responses and reproductive outcome", J Reprod Immunol, 83, pp.185~189 (2009)
- Cotch MF, "Trichomonas vaginalis associated with low birth weight and preterm delivery", Sex Transm Infect, 24, pp.353~360 (1997)
- Kigozi GG, "Treatment of Trichomonas in pregnancy and adverse outcomes of pregnancy: a subanalysis of a randomized trial in Rakai, Uganda", Am J Obstet Gynecol, 189, pp.1398~1400 (2003)
- Moodley P, "Trichomonas vaginalis is associated with pelvic inflammatory disease in women infected with human immunodeficiency virus", Clin Infect Dis, 34, pp.519~522 (2002)
- Rendón-Maldonado J, "Trichomonas vaginalis: in vitro attachment and internalization of HIV-1 and HIV-1-infected lymphocytes", J Eukaryot Microbiol, 50, pp.43~48 (2003)
- Guenthner PC, "Trichomonas vaginalis-induced epithelial monolayer disruption and human immunodeficiency virus type 1 (HIV-1) replication: implications for the sexual transmission of HIV-1", Infect Immun, 73, pp.4155~4160 (2005)
- Kissinger P, "Trichomonas vaginalis treatment reduces vaginal HIV-1 shedding", Sex Transm Infect, 36, pp.11~16 (2009)
- Workowski KA, "Sexually transmitted diseases treatment guidelines, 2010", MMWR Recomm Rep, 59, pp.1~110 (2010)
- Radonjic IV, "Diagnosis of Trichomonas vaginalis infection: The sensitivities and specificities of microscopy, culture and PCR assay", Eur J Obstet Gynecol Reprod Biol, 126, pp.116~120 (2006)
- Lawing LF, "Detection of trichomonosis in vaginal and urine specimens from women by culture and PCR", J Clin Microbiol, 38, pp.3585~3588 (2000)
- Schirm J, "Trichomonas vaginalis detection using real-time TaqMan PCR", J Microbiol Methods, 68, pp.243~247 (2007)
- Notomi T, "Loop-mediated isothermal amplification of DNA", Nucleic Acids Res, 28, pp.E63~ (2000)
- Mori Y, "Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases", J Infect Chemother, 15, pp.62~69 (2009)
- Mori Y, "Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation", Biochem Biophys Res Commun, 289, pp.150~154 (2001)
- Goto M, "Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue", Biotechniques, 46, pp.167~172 (2009)
- Tomita N, "Loop-mediated isothermal amplification(LAMP)of gene sequences and simple visual detection of products", Nat Protoc, 3, pp.877~882 (2008)
- Han ET, "Detection of four Plasmodium species by genus-and species-specific loop-mediated isothermal amplification for clinical diagnosis", J Clin Microbiol, 45, pp.2521~2528 (2007)
- Tao ZY, "Adaptation of a visualized loop-mediated isothermal amplification technique for field detection of Plasmodium vivax infection", Parasit Vectors, 4, pp.115~ (2011)
- Patel JC, "Real-time loop-mediated isothermal amplification (RealAmp) for the species-specific identification of Plasmodium vivax", PLoS One, 8, pp.e54986~ (2013)
- Karanis P, "LAMP--a powerful and flexible tool for monitoring microbial pathogens", Trends Parasitol, 25, pp.498~499 (2009)
- Crucitti T, "Comparison of culture and different PCR assays for detection of Trichomonas vaginalis in self collected vaginal swab specimens", Sex Transm Infect, 79, pp.393~398 (2003)
- Kengne P, "Trichomonas vaginalis: repeated DNA target for highly sensitive and specific polymerase chain reaction diagnosis", Cell Mol Biol(Noisy-le-grand), 40, pp.819~831 (1994)
- Lawing LF, "Detection of trichomonosis in vaginal and urine specimens from women by culture and PCR", J Clin Microbiol, 38, pp.3585~3588 (2000)
- Katiyar SK, "Beta-tubulin genes of Trichomonas vaginalis", Mol Biochem Parasitol, 64, pp.33~42 (1994)
- Madico G, "Diagnosis of Trichomonas vaginalis infection by PCR using vaginal swab samples", J Clin Microbiol, 36, pp.3205~3210 (1998)
- Crucitti T, "Molecular typing of the actin gene of Trichomonas vaginalis isolates by PCR-restriction fragment length polymorphism", Clin Microbiol Infect, 14, pp.844~852 (2008)
- Espinosa N, "Differences between coding and non-coding regions in the Trichomonas vaginalis genome: an actin gene as a locus model", Acta tropica, 78, pp.147~154 (2001)
- Notomi T, "Loop-mediated isothermal amplification of DNA", Nucleic Acids Res, 28, pp.E63~ (2000)
- 구윤경, "Prevalence of Trichomonas vaginalis in Women Visiting 2 Obstetrics and Gynecology Clinics in Daegu, South Korea", The Korean Journal of Parasitology, 54(1), pp.75~80 (2016), 대한기생충학ㆍ열대의학회
- Madico G, "Diagnosis of Trichomonas vaginalis infection by PCR using vaginal swab samples", J Clin Microbiol, 36, pp.3205~3210 (1998)
- Reyes JCB, "Development of a loop-mediated isothermal amplification assay for detection of Trichomonas vaginalis", Diagn Microbiol Infect Dis, 79, pp.337~341 (2014)
