Diagnosis of Genitourinary Tuberculosis by Loop-Mediated Isothermal Amplification Based on SYBR Green I Dye Reaction | Zendy

Ekta Kamra | Zendy; Netrapal Singh | Zendy; Anish Khan | Zendy; Jaideep Singh | Zendy; Meenakshi Chauhan | Zendy; Hemant Kamal | Zendy; Promod K. Mehta | Zendy

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Diagnosis 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

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 , biology , real time polymerase chain reaction , gene , dna , genetics

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.

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