Abstract
Human adenovirus (HAdV) is increasingly recognized as a major cause of human respiratory tract viral infections. Its outbreaks and epidemics in various populations resulted in considerable morbidity and mortality. Therefore, a rapid and specific assay for HAdV in clinical samples is of crucial importance to diagnosing HAdV infections. The present study aimed to develop and evaluate a multiplex quantitative polymerase chain reaction (qPCR) assay for the rapid detection and accurate quantification of HAdV B, C and E. The lower limit of detection for this assay was two genomic copies per reaction, and quantitative linearity ranged from 2 to 2×106 copies per reaction of the input viral DNA. Furthermore, 3,160 throat swab samples that tested HAdV negative by the immunofluorescence assay were collected and retested using the multiplex qPCR assay. The results showed that 2,906 samples were HAdV negative and the other 254 samples were HAdV positive. The HAdV species identified included B (184 samples), C (51 samples), and E (39 samples). Among the three HAdV species, HAdV B and E were detected from 8 samples, and HAdV C and E were detected from other 12 samples. The overall results demonstrated that the sensitivity and specificity of the proposed assay were 100% (254/254) and 99.6% (2894/2906), respectively. From the perspective of routine clinical diagnosis, this assay represented a rapid (≤1.5 h) and economic strategy, and had the potential to be used for the rapid and accurate diagnosis of human respiratory infections caused by HAdV B, C and E.
Highlights
Human adenoviruses (HAdVs), known as nonenveloped double‐stranded DNA viruses, are members of the Adenoviridae family with a size ranging from 70 to 100 nm [1]
HAdV serotypes that could be detected by designing primers for the hexon gene included HAdV‐C1, C2, B3, E4, C5, C6, B7, B11, B14, B16, B21, B34, and B35
HAdV serotypes that could be detected by designing primers for the fiber gene included HAdV‐C2, B3, E4, C5, B7, B11, B16, B21, B34, and B35
Summary
Human adenoviruses (HAdVs), known as nonenveloped double‐stranded DNA viruses, are members of the Adenoviridae family with a size ranging from 70 to 100 nm [1]. More than 84 genotypes, including all previously characterized serotypes, have been identified and grouped into seven different species (A‐G) (http://hadvwg.gmu.edu/), based on their immunochemical responses, nucleic acid characteristics, hexon and fiber protein characteristics, biological properties, and phylogenetic relationships [2,3,4,5,6] This subdivision has some clinical relevance, as distinct adenovirus species show a preference for specific organs: C, E, and some B species typically infect the respiratory tract; other B species infect the urinary tract; species A and F target the gastrointestinal tract; and species D target the eyes [7,8]. Researchers are constantly inventing and optimizing methods to detect adenovirus so as to avoid the overuse of antibiotics, improve the level of diagnosis and treatment, and provide a scientific basis for the prevention and control of HAdV causing respiratory illness
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