Application of dual amplification assay to identify the potentially viral pathogens of upper respiratory infection in children

Abstract

Objective To evaluate a new multiple nucleic acid detection technology-dual amplification assay, for the clinical value in diagnosis of children with upper respiratory tract infection. Methods Samples from 43 patients were tested by dual amplification assay and direct immunofluorescence assay in nasopharyngeal and throat swabs. Seven respiratory tract pathogens were analyzed, included influenzavirus A (FluA), influenzavirus B (FluB), respiratory syncytial virus (RSV), parainfluenzavirus (PIV), adenovirus (Adv), Mycoplasma pneumoniae (Mp) and Chlamydiae pneumoniae (Cpn). In addition, the samples with different result were confirmed by reverse transcription (RT)-nest PCR assay. Results The positive rate of double amplification assay were 53.3%, with 8.8% multiple infection. Excluding the detection of Mp and Cpn, the positive rate and mulitple infection rate of dual amplification was higher than direct immunofluorescence by 11.1% and 4.5% respectively. Eight samples with different result were confirmed the same as the dual amplification result by RT-nest PCR assay. Conclusions The RNA of seven pathogens can be detected simultaneously by dual amplification assay with higher sensitivity and specificity. The dual amplification technology with high clinical application value can provide comprehensive etiological diagnosis information assisting the diagnosis of upper respiratory tract infectious diseases. Key words: Upper respiratory tract infectious diseases; Pathogens; Dual amplification assay; Children