Development of a visible loop mediated isothermal amplification assay for rapid detection of Bacillus anthracis. (January 2021)

• Record Type: Journal Article
• Authors: Upadhyay, L.; Chaturvedi, V.K.; Gupta, P.K.; Sunita, S.C.; Sumithra, T.G.; Prusty, B.R.; Yadav, A.K.
• Is Part Of: Biologicals. Volume 69(2021)
• Page Start: 59
• Page End: 65
• Publication Date: 2021-01
• Subjects: Bacillus anthracis -- Pag gene -- LAMP; Biological products -- Standards -- Periodicals; Biological Products -- Periodicals; Biological Products -- standards -- Periodicals; Produits biologiques -- Normes -- Périodiques; Biological products -- Standards; Periodicals; 615.37
• DOI: 10.1016/j.biologicals.2020.11.004
• Languages: English
• ISSNs: 1045-1056

Abstract

Abstract: Distressing effects on animal and human health with lethal progression, being used as bioweapon and shared features with non-pathogenic bacteria demands sensitive, specific, safe, cost effective and rapid detection methods for anthrax causing organisms. Conventional microbiology based diagnostics for anthrax are time consuming and need sophisticated equipment, while molecular diagnostics require less time and labor. The Loop mediated isothermal amplification assay (LAMP) is rapid, sensitive and specific assay and requires no specialized equipment. In the present study, we developed a LAMP assay for rapid as well as specific detection of Bacillus anthracis . The optimized assay produced positive results with the Sterne strain and one field isolate of B. anthracis and, negative results with other bacteria of the same and different genera within 2 h. Sensitivity was 500 fg of total DNA of B. anthracis, which was 100 times more sensitive than conventional PCR. The present study also demonstrated that the simple method of total DNA extraction by repeated boiling and freezing will not adversely affect the LAMP results. In conclusion, the optimized LAMP assay is a promising tool for the specific, sensitive, less time-consuming diagnosis for anthrax causing bacteria and also, for detecting the virulence of suspected B. anthracis cultures. Highlights: Optimized a specific LAMP-PCR for B. anthracis . Sensitivity was 500 fg and 50 pg of DNA with and without loop primers respectively. Promising diagnostic tool for anthrax in an easy, time and cost-effective manner. Can also be applied for testing the virulence of suspected cultures.