Cargando…

Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus

Duck Tembusu virus (DTMUV), duck circovirus (DuCV), and new duck reovirus (NDRV) have seriously hindered the development of the poultry industry in China. To detect the three pathogens simultaneously, a multiplex digital PCR (dPCR) was developed and compared with multiplex qPCR in this study. The mu...

Descripción completa

Detalles Bibliográficos
Autores principales: Yin, Yanwen, Xiong, Chenyong, Shi, Kaichuang, Long, Feng, Feng, Shuping, Qu, Sujie, Lu, Wenjun, Huang, Meizhi, Lin, Changhua, Sun, Wenchao, Li, Zongqiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10469322/
https://www.ncbi.nlm.nih.gov/pubmed/37662994
http://dx.doi.org/10.3389/fvets.2023.1222789
_version_ 1785099415381344256
author Yin, Yanwen
Xiong, Chenyong
Shi, Kaichuang
Long, Feng
Feng, Shuping
Qu, Sujie
Lu, Wenjun
Huang, Meizhi
Lin, Changhua
Sun, Wenchao
Li, Zongqiang
author_facet Yin, Yanwen
Xiong, Chenyong
Shi, Kaichuang
Long, Feng
Feng, Shuping
Qu, Sujie
Lu, Wenjun
Huang, Meizhi
Lin, Changhua
Sun, Wenchao
Li, Zongqiang
author_sort Yin, Yanwen
collection PubMed
description Duck Tembusu virus (DTMUV), duck circovirus (DuCV), and new duck reovirus (NDRV) have seriously hindered the development of the poultry industry in China. To detect the three pathogens simultaneously, a multiplex digital PCR (dPCR) was developed and compared with multiplex qPCR in this study. The multiplex dPCR was able to specifically detect DTMUV, DuCV, and NDRV but not amplify Muscovy duck reovirus (MDRV), Muscovy duck parvovirus (MDPV), goose parvovirus (GPV), H4 avian influenza virus (H4 AIV), H6 avian influenza virus (H6 AIV), and Newcastle disease virus (NDV). The standard curves showed excellent linearity in multiplex dPCR and qPCR and were positively correlated. The sensitivity results showed that the lowest detection limit of multiplex dPCR was 1.3 copies/μL, which was 10 times higher than that of multiplex qPCR. The reproducibility results showed that the intra- and interassay coefficients of variation were 0.06–1.94%. A total of 173 clinical samples were tested to assess the usefulness of the method; the positive detection rates for DTMUV, DuCV, and NDRV were 18.5, 29.5, and 14.5%, respectively, which were approximately 4% higher than those of multiplex qPCR, and the kappa values for the clinical detection results of multiplex dPCR and qPCR were 0.85, 0.89, and 0.86, indicating that the two methods were in excellent agreement.
format Online
Article
Text
id pubmed-10469322
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-104693222023-09-01 Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus Yin, Yanwen Xiong, Chenyong Shi, Kaichuang Long, Feng Feng, Shuping Qu, Sujie Lu, Wenjun Huang, Meizhi Lin, Changhua Sun, Wenchao Li, Zongqiang Front Vet Sci Veterinary Science Duck Tembusu virus (DTMUV), duck circovirus (DuCV), and new duck reovirus (NDRV) have seriously hindered the development of the poultry industry in China. To detect the three pathogens simultaneously, a multiplex digital PCR (dPCR) was developed and compared with multiplex qPCR in this study. The multiplex dPCR was able to specifically detect DTMUV, DuCV, and NDRV but not amplify Muscovy duck reovirus (MDRV), Muscovy duck parvovirus (MDPV), goose parvovirus (GPV), H4 avian influenza virus (H4 AIV), H6 avian influenza virus (H6 AIV), and Newcastle disease virus (NDV). The standard curves showed excellent linearity in multiplex dPCR and qPCR and were positively correlated. The sensitivity results showed that the lowest detection limit of multiplex dPCR was 1.3 copies/μL, which was 10 times higher than that of multiplex qPCR. The reproducibility results showed that the intra- and interassay coefficients of variation were 0.06–1.94%. A total of 173 clinical samples were tested to assess the usefulness of the method; the positive detection rates for DTMUV, DuCV, and NDRV were 18.5, 29.5, and 14.5%, respectively, which were approximately 4% higher than those of multiplex qPCR, and the kappa values for the clinical detection results of multiplex dPCR and qPCR were 0.85, 0.89, and 0.86, indicating that the two methods were in excellent agreement. Frontiers Media S.A. 2023-08-10 /pmc/articles/PMC10469322/ /pubmed/37662994 http://dx.doi.org/10.3389/fvets.2023.1222789 Text en Copyright © 2023 Yin, Xiong, Shi, Long, Feng, Qu, Lu, Huang, Lin, Sun and Li. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Yin, Yanwen
Xiong, Chenyong
Shi, Kaichuang
Long, Feng
Feng, Shuping
Qu, Sujie
Lu, Wenjun
Huang, Meizhi
Lin, Changhua
Sun, Wenchao
Li, Zongqiang
Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus
title Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus
title_full Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus
title_fullStr Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus
title_full_unstemmed Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus
title_short Multiplex digital PCR: a superior technique to qPCR for the simultaneous detection of duck Tembusu virus, duck circovirus, and new duck reovirus
title_sort multiplex digital pcr: a superior technique to qpcr for the simultaneous detection of duck tembusu virus, duck circovirus, and new duck reovirus
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10469322/
https://www.ncbi.nlm.nih.gov/pubmed/37662994
http://dx.doi.org/10.3389/fvets.2023.1222789
work_keys_str_mv AT yinyanwen multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT xiongchenyong multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT shikaichuang multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT longfeng multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT fengshuping multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT qusujie multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT luwenjun multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT huangmeizhi multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT linchanghua multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT sunwenchao multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus
AT lizongqiang multiplexdigitalpcrasuperiortechniquetoqpcrforthesimultaneousdetectionofducktembusuvirusduckcircovirusandnewduckreovirus