Development of a loop-mediated isothermal amplification assay for detection of Trichomonas vaginalis

Reyes, John Carlo B.; Solon, Juan Antonio A.; Rivera, Windell L.

doi:10.1016/j.diagmicrobio.2014.03.016

Cited by 28 publications

(21 citation statements)

References 29 publications

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“…It is also known that DNA purification from samples could be omitted in LAMP reactions, since LAMP assays have shown a significant tolerance to inhibitor substances derived from a number of biological samples [ 71 ], [ 72 ], [ 73 ]. Additionally, other LAMP assays with high sensitivity and no complicate requirement procedure for DNA extraction have been developed for molecular detection and diagnostic of bacterial [ 74 ] and parasitic [ 75 ] diseases in urine samples. Moreover, a simple heating DNA obtaining method has been also successfully applied with other clinical samples, such us blood [ 41 ] and swaps [ 42 ] in LAMP amplification of both Plasmodium and Leishmania species nucleic acids, respectively.…”

Section: Discussionmentioning

confidence: 99%

The Rapid-Heat LAMPellet Method: A Potential Diagnostic Method for Human Urogenital Schistosomiasis

et al. 2015

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BackgroundUrogenital schistosomiasis due to Schistosoma haematobium is a serious underestimated public health problem affecting 112 million people - particularly in sub-Saharan Africa. Microscopic examination of urine samples to detect parasite eggs still remains as definitive diagnosis. This work was focussed on developing a novel loop-mediated isothermal amplification (LAMP) assay for detection of S. haematobium DNA in human urine samples as a high-throughput, simple, accurate and affordable diagnostic tool to use in diagnosis of urogenital schistosomiasis.Methodology/Principal FindingsA LAMP assay targeting a species specific sequence of S. haematobium ribosomal intergenic spacer was designed. The effectiveness of our LAMP was assessed in a number of patients´ urine samples with microscopy confirmed S. haematobium infection. For potentially large-scale application in field conditions, different DNA extraction methods, including a commercial kit, a modified NaOH extraction method and a rapid heating method were tested using small volumes of urine fractions (whole urine, supernatants and pellets). The heating of pellets from clinical samples was the most efficient method to obtain good-quality DNA detectable by LAMP. The detection limit of our LAMP was 1 fg/µL of S. haematobium DNA in urine samples. When testing all patients´ urine samples included in our study, diagnostic parameters for sensitivity and specificity were calculated for LAMP assay, 100% sensitivity (95% CI: 81.32%-100%) and 86.67% specificity (95% CI: 75.40%-94.05%), and also for microscopy detection of eggs in urine samples, 69.23% sensitivity (95% CI: 48.21% -85.63%) and 100% specificity (95% CI: 93.08%-100%).Conclusions/SignificanceWe have developed and evaluated, for the first time, a LAMP assay for detection of S. haematobium DNA in heated pellets from patients´ urine samples using no complicated requirement procedure for DNA extraction. The procedure has been named the Rapid-Heat LAMPellet method and has the potential to be developed further as a field diagnostic tool for use in urogenital schistosomiasis-endemic areas.

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Section: Discussionmentioning

confidence: 99%

The Rapid-Heat LAMPellet Method: A Potential Diagnostic Method for Human Urogenital Schistosomiasis

et al. 2015

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“…PCR provides sensitivities of 88-97%, depending on the specimen type, population and reference standard (Sherrard et al, 2014). A LAMP assay targeting a 2-kbp repeated sequence has also been described and demonstrated 10-1000 times higher sensitivity than the comparator 18S rDNA PCR (Reyes, Solon, & Rivera, 2014), with an LoD of 1 trichomonad per reaction. These NAATs can be used to test a variety of specimen types including vaginal and urethral swabs, urine and semen.…”

Section: Diagnostic Modalitiesmentioning

confidence: 97%

Molecular Diagnostics in the Diagnosis of Parasitic Infection

Pritt¹

2015

Methods in Microbiology

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“…Finally, recent proof-of-concept studies have discussed detection of T. vaginalis by loop-mediated isothermal amplification 61 and PCR-based microarray. 62…”

Section: Extra-urogenital Detectionmentioning

confidence: 99%

Update on Laboratory Diagnosis and Epidemiology of Trichomonas vaginalis: You Can Teach an “Old” Dog “New” Trichs

Munson

Napierala

Munson

2016

Clinical Microbiology Newsletter

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Past viewpoints on Trichomonas vaginalis infection have characterized the associated clinical disease as a "nuisance" condition, with affected demographics largely being older African American females residing in urban centers. The advent of commercial molecular assays specific for T. vaginalishas offered a new outlook on trichomoniasis. Within high-prevalence sexually transmitted infection populations, parasite distribution is not localized to specific population centers, and T. vaginalis prevalence is elevated among both younger and older age groups. Adaptation of these molecular assays can additionally facilitate male screening and subsequent epidemiologic characterization. These findings, combined with associations between T. vaginalis infection and human immunodeficiency virus (HIV) acquisition/transmission and persistent human papillomavirus infection, support consideration of the expansion of T. vaginalis screening efforts in the realms of clinical practice and public health.

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Development of a loop-mediated isothermal amplification assay for detection of Trichomonas vaginalis

Cited by 28 publications

References 29 publications

The Rapid-Heat LAMPellet Method: A Potential Diagnostic Method for Human Urogenital Schistosomiasis

The Rapid-Heat LAMPellet Method: A Potential Diagnostic Method for Human Urogenital Schistosomiasis

Molecular Diagnostics in the Diagnosis of Parasitic Infection

Update on Laboratory Diagnosis and Epidemiology of Trichomonas vaginalis: You Can Teach an “Old” Dog “New” Trichs

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