Abstract
BackgroundRespiratory infections are a serious threat to human health. So, rapid detection of all respiratory pathogens can facilitate prompt treatment and prevent the deterioration of respiratory disease. Previously published primers and probes of the TaqMan array card (TAC) for respiratory pathogens are not sensitive to Chinese clinical specimens. This study aimed to develop and improve the TAC assay to detect 28 respiratory viral and bacterial pathogens in a Chinese population.MethodsTo improve the sensitivity, we redesigned the primers and probes, and labeled the probes with minor groove binders. The amplification efficiency, sensitivity, and specificity of the primers and probes were determined using target-gene containing standard plasmids. The detection performance of the TAC was evaluated on 754 clinical specimens and the results were compared with those from conventional methods.ResultsThe performance of the TAC assay was evaluated using 754 clinical throat swab samples and the results were compared with those from gold-standard methods. The sensitivity and specificity were 95.4 and 96.6%, respectively. The lowest detection limit of the TAC was 10 to 100 copies/μL.ConclusionsTAC is an efficient, accurate, and high-throughput approach to detecting multiple respiratory pathogens simultaneously and is a promising tool for the identification of pathogen outbreaks.
Highlights
Respiratory infections are a serious threat to human health
Strains Bacterial and viral strains that had been isolated from clinical specimens were used to validate the TaqMan array card (TAC)
The following 20 viruses and 8 bacteria were collected from other laboratories: influenza type A virus, influenza type B virus, enterovirus, parainfluenza virus, respiratory syncytial virus, human metapneumovirus, adenovirus, rhinovirus, human bocavirus, human coronavirus
Summary
Respiratory infections are a serious threat to human health. So, rapid detection of all respiratory pathogens can facilitate prompt treatment and prevent the deterioration of respiratory disease. This study aimed to develop and improve the TAC assay to detect 28 respiratory viral and bacterial pathogens in a Chinese population. Respiratory diseases, such as acute respiratory infections (ARIs) and community-acquired pneumonia (CAP), are a serious threat to human health [1,2,3,4,5,6,7]. Serology, ELISA, immunofluorescence staining, and conventional molecular diagnostics such as PCR and RT-PCR are common conventional approaches to detecting respiratory pathogens These conventional detection methods have disadvantages; they have low sensitivity and are time-consuming, laborintensive and susceptible to contamination [1, 3, 14]. The real-time PCR has been gradually applied to clinical
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