A fluorescent bead-based multiplex PCR assay for the simultaneous detection of seven viral diseases endemic in Africa / 中华实验和临床病毒学杂志

ABSTRACT

Objective@#To establish a fluorescent bead-based multiplex assay for the simultaneous detection of seven viral diseases endemic in Africa.@*Methods@#The genomic sequences of the viral pathogens causing Rift valley fever, Yellow fever, Marburg virus disease, Ebola virus disease, Lassa fever, Crimean-Congo hemorrhagic fever and Chikungunya fever were compared, PCR detection target fragments were selected, and amplification primers and hybrid probes were designed. The reference samples of related pathogens were prepared by chemical synthesis of DNA and in vitro transcription RNA. The sensitivity and stability of the detection method were evaluated. The specificity was evaluated by testing 30 samples of suspected dengue fever, and hantavirus diseases, and 32 healthy human blood samples.@*Results@#The fluorescent bead-based multiplex assay could specifically detect the corresponding pathogen, the detection limit was at a range of 102-105 copies/ μl, the specificity was 100%, and the intra-assay coefficient of variation was below 12%, and the inter-assay coefficient of variation was below 15%.@*Conclusions@#A fluorescent bead-based multiplex PCR assay for the simultaneous detection of seven viral diseases endemic in Africa was established, which may provide a new choice for the screening of suspected infectious diseases.