A novel fluoro colorimetric Loop mediated isothermal amplification (LAMP) assay for detection of Trichomonasvaginalis.
Indian J Med Microbiol
; 49: 100610, 2024.
Article
in En
| MEDLINE
| ID: mdl-38735643
ABSTRACT
BACKGROUND:
Trichomoniasis is a curable, non-viral, sexually transmitted infection. Early diagnosis and treatment of cases can prevent complications and further spread of infection. Rapid diagnostics tests, which can be performed on-site, will help in early diagnosis. The study aims to develop a rapid diagnostic test based on the principle of fluoro-colorimetric LAMP for detecting Trichomonas vaginalis (TV). MATERIALS ANDMETHODS:
T. vaginalis was grown in a modified CPLM medium, and DNA was extracted. Three pairs of LAMP primers targeting the actin gene were designed using the primerexplorer V.5 online tool. The LAMP assay was standardized for temperature and time. To determine the LAMP assay's detection limit, diluted TV DNA and spiked urine samples were used. Conventional PCR was done using previously published primers and compared with LAMP results. The sensitivity and specificity to detect TV from clinical specimens were assessed.RESULTS:
The optimum performance of the LAMP assay was determined to be 63 °C for 60 min and terminated at 80 °C for 5 min. The LAMP assay could detect 60 fg/µl of diluted TV DNA and up to 1 parasite/ml of spiked samples. The assay was 1000 times more sensitive than PCR. The LAMP assay was 100% sensitive and specific with crude extract, and reactions were visually discernible. INTERPRETATION &CONCLUSIONS:
The LAMP assay developed in the study is easy to perform and interpret, affordable, rapid, and highly sensitive to detect T. vaginalis. It is ideally suited for the point-of-care test, as it fulfills WHO's recommended ASSURED characteristics.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Trichomonas vaginalis
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Sensitivity and Specificity
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Colorimetry
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Nucleic Acid Amplification Techniques
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Molecular Diagnostic Techniques
Limits:
Female
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Humans
Language:
En
Journal:
Indian J Med Microbiol
/
Indian j. med. microbiol.
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Indian journal of medical microbiology
Journal subject:
MICROBIOLOGIA
Year:
2024
Document type:
Article
Country of publication: