The use of loop-mediated isothermal DNA amplification for the detection and identification of the anthrax pathogen

Abstract

⎯The results of detection and identification of Bacillus anthracis strains in loop-mediated isothermal DNA amplification (LAMP) reaction performed under optimized conditions with original primers and thermostable DNA polymerase are presented. Reproducible LAMP-based detection of chromosomal and plasmid DNA targets specific for B. anthracis strains has been demonstrated. No cross reactions with DNA from bacterial strains of other species of the B. cereus group were detected. The development of tests for anthrax-pathogen detection based on the optimized reaction of loop isothermal DNA amplification is planned. These tests will be convenient for clinical studies and field diagnostics due to the absence of requirements for sophisticated equipment.

Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA

“…SD DNA polymerase used to amplify the cDNA is a thermostable Taq DNA polymerase mutant that has strong 5′–3′ strand displacement and 5′–3′ polymerase activities. This polymerase is suitable for PCR, PCDR, and isothermal DNA amplifications ( Ignatov et al, 2014 ; Shchit et al, 2017 ; Smith, 2017 ; Alyethodi et al, 2018 ; Lou et al, 2018 ; Wang et al, 2018 ). The mixes for performing either qPCR or qPCDR were identical, except for two extra outer primers added in case of qPCDR.…”

Combinations of PCR and isothermal amplification techniques are suitable for fast and sensitive detection of SARS-CoV-2 viral RNA

“…SD DNA polymerase used to amplify the cDNA is a thermostable Taq DNA polymerase mutant that has strong 5’–3’ strand displacement and 5’–3’ polymerase activities. This polymerase is suitable for PCR, PCDR, and isothermal DNA amplifications (Wang et al, 2018; Shchit et al, 2017; Ignatov et al, 2014; Smith, 2017; Alyethodi et al, 2018; Lou et al, 2018). The mixes for performing either qPCR or qPCDR were identical, except for two extra outer primers added in case of qPCDR.…”

Isothermal Nucleic Acid Amplification Techniques and Their Use in Bioanalysis

“…LAMP is widely used as a screening technique, because it can be conducted at a constant temperature and is highly efficient and specific. LAMP has been used for identification of Mycobacterium tuberculosis, herpes, severe acute respiratory syndrome, anthrax, human and avian influenza viruses, and other pathogens [12,13]. LAMP was able to detect Leptospira DNA at a concen tration as low as 200 pg/ml.…”