KI and WU are two recently-described human polyomaviruses worldwide distributed. An association between these two viruses and respiratory infections has been suggested, in particular in children and immunocompromised patients. Since no standardized detection methods are currently available, we developed a duplex real-time PCR assay for the simultaneous detection of KIHPyV and WUHPyV polyomaviruses based on TaqMan probes. We assessed this technique on 831 nasopharyngeal aspirate specimens from hospitalized paediatric patients with respiratory symptoms, retrospectively analysed with commercial multiplex assay for 16 other major respiratory viruses. Our assay detected 500 copies/mL for both KIHPyV and WUHPyV in all tested samples. We detected KIHPyV and WUHPyV genome in 28 (3.36%) and 41 samples (4.93%) respectively. Moreover, in 3 samples, the co-infection of the two viruses was found. 55 out of all positive samples (n=69) for KIHPyV and/or WUHPyV infection exhibited a co-infection with one or more respiratory viruses, confirming that KIHPyV and WUHPyV were often detected in association with other viral infections. Interestingly, KIHPyV and WUHPyV were detected singularly in 8 out of 28 cases and 6 out of 41 cases, respectively, suggesting a possible direct role of these viruses in the respiratory diseases. In conclusion, this method could be taken into account as an alternative technical approach to detect KIHPyV and/or WUHPyV in respiratory samples helping in the definition of the pathogenic potential and of the epidemiological prevalence of these two viruses.
Development of a new duplex real-time polymerase chain reaction assay for detection and quantification of KIHPyV and WUHPyV in pediatric samples
CARELLI, MARIA
2019-01-01
Abstract
KI and WU are two recently-described human polyomaviruses worldwide distributed. An association between these two viruses and respiratory infections has been suggested, in particular in children and immunocompromised patients. Since no standardized detection methods are currently available, we developed a duplex real-time PCR assay for the simultaneous detection of KIHPyV and WUHPyV polyomaviruses based on TaqMan probes. We assessed this technique on 831 nasopharyngeal aspirate specimens from hospitalized paediatric patients with respiratory symptoms, retrospectively analysed with commercial multiplex assay for 16 other major respiratory viruses. Our assay detected 500 copies/mL for both KIHPyV and WUHPyV in all tested samples. We detected KIHPyV and WUHPyV genome in 28 (3.36%) and 41 samples (4.93%) respectively. Moreover, in 3 samples, the co-infection of the two viruses was found. 55 out of all positive samples (n=69) for KIHPyV and/or WUHPyV infection exhibited a co-infection with one or more respiratory viruses, confirming that KIHPyV and WUHPyV were often detected in association with other viral infections. Interestingly, KIHPyV and WUHPyV were detected singularly in 8 out of 28 cases and 6 out of 41 cases, respectively, suggesting a possible direct role of these viruses in the respiratory diseases. In conclusion, this method could be taken into account as an alternative technical approach to detect KIHPyV and/or WUHPyV in respiratory samples helping in the definition of the pathogenic potential and of the epidemiological prevalence of these two viruses.File | Dimensione | Formato | |
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DEVELOPMENT OF A NEW DUPLEX REAL-TIME POLYMERASE CHAIN REACTION ASSAY FOR DETECTION AND QUANTIFICATION OF .pdf
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