Open AccessDiagnosis of genitourinary tuberculosis by loop-mediated isothermal amplification based on SYBR Green I dye reaction
Author(s) -
Ekta Kamra,
Netrapal Singh,
Anish Khan,
Jaideep Singh,
Meenakshi Chauhan,
Hemant Kamal,
Promod K. Mehta
Publication year - 2022
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2022-0027
Subject(s) - loop mediated isothermal amplification , sybr green i , multiplex , microbiology and biotechnology , real time polymerase chain reaction , biology , gene , genetics , dna
A multitargeted loop-mediated isothermal amplification (MT-LAMP) assay targeting mpt64 (Rv1980c) and IS 6110 was designed to diagnose genitourinary tuberculosis (GUTB) cases. While assessing gel-based, hydroxynaphthol blue (HNB) and SYBR Green I MT-LAMP assays on GUTB specimens (n = 28) in a pilot study, both gel-based/SYBR Green I assays exhibited better sensitivity than HNB LAMP. Since SYBR Green MT-LAMP is easier to perform compared with a gel-based assay, a higher number of GUTB specimens (n = 55) were evaluated by SYBR Green MT-LAMP, wherein 85.5% sensitivity and 94.4% specificity (n = 36) were obtained. Moreover, the sensitivity attained by MT-LAMP was significantly higher (p < 0.05) than with multiplex-PCR ( mpt64 + IS 6110 ). After further validating these MT-LAMP data in different epidemiological settings, this assay may be developed as a diagnostic kit.