Producing positive control materials for template in PCR testing to detect botulinum neurotoxin types A and B genes

Abstract

Botulinum neurotoxin (BoNT) is produced by the bacterium Clostridium botulinum and some other bacterial strains such as C. butyricum and C. baratii. It is considered the strongest toxin known to humans and can cause botulism. Botulinum neurotoxin can be fatal in humans and most botulism cases are caused by type A and B toxins. In this study, the research team used C. botulinum strains isolated from botulism poisoning cases in Vietnam, which were identified to produce type A and B toxins, from which we successfully transformed plasmids carrying genes specific for BoNT type A and type B genes based on the reference gene sequence according to TCVN 11395:2016. These plasmids were successfully used as template DNA for PCR reactions. In the PCR reaction, plasmids used to transform genes producing botulinum toxin types A and B have a detection limit of 102 copies/µL. The study has fully verified the validation parameters, with 100% accuracy and specificity.