The Application of Multiplex PCR-MALDI-TOF MS Technique for Identification of Omicron EG.5 and BA.2.86 Variants
In light of the highly transmissible nature of the Omicron EG.5 and BA.2.86 variants,this study integrates Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry(MALDI-TOF MS)with multiplex PCR technology.Primers were designed to amplify and extend key mutation sites on the spike(S)protein of these variants.Additionally,the D614G mutation site was included for identification,along with the ORF1ab and N gene detection sites for the identification of SARS-CoV-2,enabling the simultaneous detection of 10 characteristic sites.The results demonstrate that this method can detect both the EG.5 and BA.2.86 variants in the same reaction system with high accuracy and a sensitivity as low as 20 copies per reaction.This method is well-suited for dynamic screening of a large number of samples by customs and other epidemic monitoring departments,focusing on variants of concern.It serves as a valuable technical complement to quantitative fluorescent PCR and high-throughput sequencing methods,providing technical support for virus tracing and epidemic surveillance.