Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy

Background: Acute encephalitis is a serious neurological condition having a high mortality rate and affecting both children and adults. This study aimed to develop a multiplex PCR method for the simultaneous screening of clinical samples for the presence of the 10 viruses presently considered as the...

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Main Authors: Ngan T.K. Pham, Hiroshi Ushijima, Aksara Thongprachum, Quang D. Trinh, Pattara Khamrin, Chikako Arakawa, Wakako Ishii, Shoko Okitsu, Shihoko Komine-Aizawa, Satoshi Hayakawa
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Published: 2018
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/46456
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-464562018-04-25T07:32:11Z Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy Ngan T.K. Pham Hiroshi Ushijima Aksara Thongprachum Quang D. Trinh Pattara Khamrin Chikako Arakawa Wakako Ishii Shoko Okitsu Shihoko Komine-Aizawa Satoshi Hayakawa Agricultural and Biological Sciences Background: Acute encephalitis is a serious neurological condition having a high mortality rate and affecting both children and adults. This study aimed to develop a multiplex PCR method for the simultaneous screening of clinical samples for the presence of the 10 viruses presently considered as the major viral causes of acute encephalitis/encephalopathy in Asia. Methods: Using previously published primers that have been widely used to screen for herpes virus-6, influenza A virus, human parechovirus, herpes simplex viruses 1 and 2, Japanese encephalitis virus, group A rotavirus, enterovirus, adenovirus, and dengue virus in clinical samples, a single-tube multiplex PCR assay was developed and was tested for its sensitivity and specificity. The method was then applied to screen 57 clinical samples, consisting of 13 fecal samples, 5 throat swabs, 3 post-nasal swabs, 18 serum samples, and 18 cerebrospinal fluid (CSF) samples, collected from 18 hospitalized Japanese children with suspected viral encephalitis/encephalopathy for the target viruses, and the results were compared with those of a monoplex PCR method. Results: Positive viral controls of the 10 viruses were correctly typed using this multiplex PCR method. The multiplex PCR method showed high specificity with no unspecific amplification to non-target viruses. The results of applying this PCR method for screening clinical samples showed that 6 fecal samples, 2 serum samples, and 1 CSF sample collected from 7 patients were positive for a virus, specifically group A rotavirus (4 patients, 22.2%), enterovirus (2 patients, 11.1%), or adenovirus (1 patient, 5.6%). In comparison with monoplex PCR, for group A rotavirus, enterovirus, and adenovirus, the sensitivity of this multiplex PCR method decreased for serum, cerebrospinal fluid, and throat swab samples. Conclusions: This newly developed multiplex PCR method is a simple, rapid diagnostic tool and can be used to screen clinical samples for viruses causing acute encephalitis/encephalopathy in children in Asian countries. 2018-04-25T06:55:08Z 2018-04-25T06:55:08Z 2017-01-01 Journal 14336510 2-s2.0-85013788666 10.7754/Clin.Lab.2016.160630 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85013788666&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/46456
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Agricultural and Biological Sciences
spellingShingle Agricultural and Biological Sciences
Ngan T.K. Pham
Hiroshi Ushijima
Aksara Thongprachum
Quang D. Trinh
Pattara Khamrin
Chikako Arakawa
Wakako Ishii
Shoko Okitsu
Shihoko Komine-Aizawa
Satoshi Hayakawa
Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy
description Background: Acute encephalitis is a serious neurological condition having a high mortality rate and affecting both children and adults. This study aimed to develop a multiplex PCR method for the simultaneous screening of clinical samples for the presence of the 10 viruses presently considered as the major viral causes of acute encephalitis/encephalopathy in Asia. Methods: Using previously published primers that have been widely used to screen for herpes virus-6, influenza A virus, human parechovirus, herpes simplex viruses 1 and 2, Japanese encephalitis virus, group A rotavirus, enterovirus, adenovirus, and dengue virus in clinical samples, a single-tube multiplex PCR assay was developed and was tested for its sensitivity and specificity. The method was then applied to screen 57 clinical samples, consisting of 13 fecal samples, 5 throat swabs, 3 post-nasal swabs, 18 serum samples, and 18 cerebrospinal fluid (CSF) samples, collected from 18 hospitalized Japanese children with suspected viral encephalitis/encephalopathy for the target viruses, and the results were compared with those of a monoplex PCR method. Results: Positive viral controls of the 10 viruses were correctly typed using this multiplex PCR method. The multiplex PCR method showed high specificity with no unspecific amplification to non-target viruses. The results of applying this PCR method for screening clinical samples showed that 6 fecal samples, 2 serum samples, and 1 CSF sample collected from 7 patients were positive for a virus, specifically group A rotavirus (4 patients, 22.2%), enterovirus (2 patients, 11.1%), or adenovirus (1 patient, 5.6%). In comparison with monoplex PCR, for group A rotavirus, enterovirus, and adenovirus, the sensitivity of this multiplex PCR method decreased for serum, cerebrospinal fluid, and throat swab samples. Conclusions: This newly developed multiplex PCR method is a simple, rapid diagnostic tool and can be used to screen clinical samples for viruses causing acute encephalitis/encephalopathy in children in Asian countries.
format Journal
author Ngan T.K. Pham
Hiroshi Ushijima
Aksara Thongprachum
Quang D. Trinh
Pattara Khamrin
Chikako Arakawa
Wakako Ishii
Shoko Okitsu
Shihoko Komine-Aizawa
Satoshi Hayakawa
author_facet Ngan T.K. Pham
Hiroshi Ushijima
Aksara Thongprachum
Quang D. Trinh
Pattara Khamrin
Chikako Arakawa
Wakako Ishii
Shoko Okitsu
Shihoko Komine-Aizawa
Satoshi Hayakawa
author_sort Ngan T.K. Pham
title Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy
title_short Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy
title_full Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy
title_fullStr Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy
title_full_unstemmed Multiplex PCR for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from Japanese children with suspected viral encephalitis/encephalopathy
title_sort multiplex pcr for the detection of 10 viruses causing encephalitis/encephalopathy and its application to clinical samples collected from japanese children with suspected viral encephalitis/encephalopathy
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85013788666&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/46456
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