Molecular strategy for the direct detection and identification of human enteroviruses in clinical specimens associated with hand, foot and mouth disease
Diseases caused by human enteroviruses (EVs) are a major global public health problem. Thus, the effective diagnosis of all human EVs infections and the monitoring of epidemiological and ecological dynamic changes are urgently needed. Based on two comprehensive virological surveillance systems of ha...
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| Veröffentlicht in: | PloS one 2020-11, Vol.15 (11), p.e0241614-e0241614 |
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| creator | Zhou, Yonghong Qiu, Qi Luo, Kaiwei Liao, Qiaohong Li, Yu Cui, Peng Liang, Lu Cheng, Yibing Wang, Lili Wang, Kai Van Tan, Le Rogier van Doorn, H Yu, Hongjie |
| description | Diseases caused by human enteroviruses (EVs) are a major global public health problem. Thus, the effective diagnosis of all human EVs infections and the monitoring of epidemiological and ecological dynamic changes are urgently needed.
Based on two comprehensive virological surveillance systems of hand, foot and mouth disease (HFMD), real-time PCR and nested RT-PCR (RT-snPCR) methods based on the enteroviral VP1, VP4-VP2 and VP4 regions were designed to directly detect all human EVs serotypes in clinical specimens.
The results showed that the proposed serotyping strategy exhibit very high diagnostic efficiency (Study 1: 99.9%; Study 2: 89.5%), and the variance between the study was due to inclusion of the specific Coxsackie virus A6 (CVA6) real-time RT-PCR and VP4 RT-snPCR in Study 1 but not Study 2. Furthermore, only throat swabs were collected and analyzed in Study 2, whereas in Study 1, if a specific EV serotype was not identified in the primary stool sample, other sample types (rectal swab and throat swab) were further tested where available. During the study period from 2013 to 2018, CVA6 became one of the main HFMD causative agents, whereas the level of enterovirus A71 (EV-A71) declined in 2017.
The findings of this study demonstrate the appropriate application of PCR methods and the combination of biological sample types that are useful for etiological studies and propose a molecular strategy for the direct detection of human EVs in clinical specimens associated with HFMD. |
| doi_str_mv | 10.1371/journal.pone.0241614 |
| format | Article |
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Based on two comprehensive virological surveillance systems of hand, foot and mouth disease (HFMD), real-time PCR and nested RT-PCR (RT-snPCR) methods based on the enteroviral VP1, VP4-VP2 and VP4 regions were designed to directly detect all human EVs serotypes in clinical specimens.
The results showed that the proposed serotyping strategy exhibit very high diagnostic efficiency (Study 1: 99.9%; Study 2: 89.5%), and the variance between the study was due to inclusion of the specific Coxsackie virus A6 (CVA6) real-time RT-PCR and VP4 RT-snPCR in Study 1 but not Study 2. Furthermore, only throat swabs were collected and analyzed in Study 2, whereas in Study 1, if a specific EV serotype was not identified in the primary stool sample, other sample types (rectal swab and throat swab) were further tested where available. During the study period from 2013 to 2018, CVA6 became one of the main HFMD causative agents, whereas the level of enterovirus A71 (EV-A71) declined in 2017.
The findings of this study demonstrate the appropriate application of PCR methods and the combination of biological sample types that are useful for etiological studies and propose a molecular strategy for the direct detection of human EVs in clinical specimens associated with HFMD.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0241614</identifier><identifier>PMID: 33166321</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adolescent ; Biological samples ; Biology and Life Sciences ; Capsid Proteins - genetics ; Causes of ; Child ; Child, Preschool ; Coxsackievirus infections ; Cytomegalovirus ; Diagnostic systems ; Disease control ; Disease prevention ; Education ; Enterovirus A, Human - genetics ; Enterovirus A, Human - isolation & purification ; Enteroviruses ; Epidemiology ; Etiology ; Feces - virology ; Hand, Foot and Mouth Disease - diagnosis ; Hand, Foot and Mouth Disease - virology ; Hand-foot-and-mouth disease ; Health surveillance ; Hospitals ; Humans ; Identification and classification ; Infant ; Infections ; Infectious diseases ; Intestinal Mucosa - virology ; Laboratories ; Medical diagnosis ; Medicine and Health Sciences ; Molecular diagnostic techniques ; Molecular Diagnostic Techniques - methods ; Molecular Diagnostic Techniques - standards ; People and Places ; Polymerase chain reaction ; Public health ; Real time ; Real-Time Polymerase Chain Reaction - methods ; Real-Time Polymerase Chain Reaction - standards ; Rectum ; Research and Analysis Methods ; Respiratory Mucosa - virology ; Reverse Transcriptase Polymerase Chain Reaction - methods ; Reverse Transcriptase Polymerase Chain Reaction - standards ; Sensitivity and Specificity ; Serotypes ; Serotyping ; Strategy ; Surveillance systems ; Testing ; Viral Structural Proteins - genetics ; Viruses ; VP1 protein</subject><ispartof>PloS one, 2020-11, Vol.15 (11), p.e0241614-e0241614</ispartof><rights>COPYRIGHT 2020 Public Library of Science</rights><rights>2020 Zhou et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2020 Zhou et al 2020 Zhou et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c653t-3b74f8c629c71f27d86863dd306fe40f72a2e8b08403206eeccc799632be8d6f3</citedby><cites>FETCH-LOGICAL-c653t-3b74f8c629c71f27d86863dd306fe40f72a2e8b08403206eeccc799632be8d6f3</cites><orcidid>0000-0002-6335-5648 ; 0000-0002-9807-1821</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7652283/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7652283/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79569,79570</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33166321$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhou, Yonghong</creatorcontrib><creatorcontrib>Qiu, Qi</creatorcontrib><creatorcontrib>Luo, Kaiwei</creatorcontrib><creatorcontrib>Liao, Qiaohong</creatorcontrib><creatorcontrib>Li, Yu</creatorcontrib><creatorcontrib>Cui, Peng</creatorcontrib><creatorcontrib>Liang, Lu</creatorcontrib><creatorcontrib>Cheng, Yibing</creatorcontrib><creatorcontrib>Wang, Lili</creatorcontrib><creatorcontrib>Wang, Kai</creatorcontrib><creatorcontrib>Van Tan, Le</creatorcontrib><creatorcontrib>Rogier van Doorn, H</creatorcontrib><creatorcontrib>Yu, Hongjie</creatorcontrib><title>Molecular strategy for the direct detection and identification of human enteroviruses in clinical specimens associated with hand, foot and mouth disease</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Diseases caused by human enteroviruses (EVs) are a major global public health problem. Thus, the effective diagnosis of all human EVs infections and the monitoring of epidemiological and ecological dynamic changes are urgently needed.
Based on two comprehensive virological surveillance systems of hand, foot and mouth disease (HFMD), real-time PCR and nested RT-PCR (RT-snPCR) methods based on the enteroviral VP1, VP4-VP2 and VP4 regions were designed to directly detect all human EVs serotypes in clinical specimens.
The results showed that the proposed serotyping strategy exhibit very high diagnostic efficiency (Study 1: 99.9%; Study 2: 89.5%), and the variance between the study was due to inclusion of the specific Coxsackie virus A6 (CVA6) real-time RT-PCR and VP4 RT-snPCR in Study 1 but not Study 2. Furthermore, only throat swabs were collected and analyzed in Study 2, whereas in Study 1, if a specific EV serotype was not identified in the primary stool sample, other sample types (rectal swab and throat swab) were further tested where available. During the study period from 2013 to 2018, CVA6 became one of the main HFMD causative agents, whereas the level of enterovirus A71 (EV-A71) declined in 2017.
The findings of this study demonstrate the appropriate application of PCR methods and the combination of biological sample types that are useful for etiological studies and propose a molecular strategy for the direct detection of human EVs in clinical specimens associated with HFMD.</description><subject>Adolescent</subject><subject>Biological samples</subject><subject>Biology and Life Sciences</subject><subject>Capsid Proteins - genetics</subject><subject>Causes of</subject><subject>Child</subject><subject>Child, Preschool</subject><subject>Coxsackievirus infections</subject><subject>Cytomegalovirus</subject><subject>Diagnostic systems</subject><subject>Disease control</subject><subject>Disease prevention</subject><subject>Education</subject><subject>Enterovirus A, Human - genetics</subject><subject>Enterovirus A, Human - isolation & purification</subject><subject>Enteroviruses</subject><subject>Epidemiology</subject><subject>Etiology</subject><subject>Feces - virology</subject><subject>Hand, Foot and Mouth Disease - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhou, Yonghong</au><au>Qiu, Qi</au><au>Luo, Kaiwei</au><au>Liao, Qiaohong</au><au>Li, Yu</au><au>Cui, Peng</au><au>Liang, Lu</au><au>Cheng, Yibing</au><au>Wang, Lili</au><au>Wang, Kai</au><au>Van Tan, Le</au><au>Rogier van Doorn, H</au><au>Yu, Hongjie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular strategy for the direct detection and identification of human enteroviruses in clinical specimens associated with hand, foot and mouth disease</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2020-11-09</date><risdate>2020</risdate><volume>15</volume><issue>11</issue><spage>e0241614</spage><epage>e0241614</epage><pages>e0241614-e0241614</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Diseases caused by human enteroviruses (EVs) are a major global public health problem. Thus, the effective diagnosis of all human EVs infections and the monitoring of epidemiological and ecological dynamic changes are urgently needed.
Based on two comprehensive virological surveillance systems of hand, foot and mouth disease (HFMD), real-time PCR and nested RT-PCR (RT-snPCR) methods based on the enteroviral VP1, VP4-VP2 and VP4 regions were designed to directly detect all human EVs serotypes in clinical specimens.
The results showed that the proposed serotyping strategy exhibit very high diagnostic efficiency (Study 1: 99.9%; Study 2: 89.5%), and the variance between the study was due to inclusion of the specific Coxsackie virus A6 (CVA6) real-time RT-PCR and VP4 RT-snPCR in Study 1 but not Study 2. Furthermore, only throat swabs were collected and analyzed in Study 2, whereas in Study 1, if a specific EV serotype was not identified in the primary stool sample, other sample types (rectal swab and throat swab) were further tested where available. During the study period from 2013 to 2018, CVA6 became one of the main HFMD causative agents, whereas the level of enterovirus A71 (EV-A71) declined in 2017.
The findings of this study demonstrate the appropriate application of PCR methods and the combination of biological sample types that are useful for etiological studies and propose a molecular strategy for the direct detection of human EVs in clinical specimens associated with HFMD.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>33166321</pmid><doi>10.1371/journal.pone.0241614</doi><tpages>e0241614</tpages><orcidid>https://orcid.org/0000-0002-6335-5648</orcidid><orcidid>https://orcid.org/0000-0002-9807-1821</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2020-11, Vol.15 (11), p.e0241614-e0241614 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2458949546 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Adolescent Biological samples Biology and Life Sciences Capsid Proteins - genetics Causes of Child Child, Preschool Coxsackievirus infections Cytomegalovirus Diagnostic systems Disease control Disease prevention Education Enterovirus A, Human - genetics Enterovirus A, Human - isolation & purification Enteroviruses Epidemiology Etiology Feces - virology Hand, Foot and Mouth Disease - diagnosis Hand, Foot and Mouth Disease - virology Hand-foot-and-mouth disease Health surveillance Hospitals Humans Identification and classification Infant Infections Infectious diseases Intestinal Mucosa - virology Laboratories Medical diagnosis Medicine and Health Sciences Molecular diagnostic techniques Molecular Diagnostic Techniques - methods Molecular Diagnostic Techniques - standards People and Places Polymerase chain reaction Public health Real time Real-Time Polymerase Chain Reaction - methods Real-Time Polymerase Chain Reaction - standards Rectum Research and Analysis Methods Respiratory Mucosa - virology Reverse Transcriptase Polymerase Chain Reaction - methods Reverse Transcriptase Polymerase Chain Reaction - standards Sensitivity and Specificity Serotypes Serotyping Strategy Surveillance systems Testing Viral Structural Proteins - genetics Viruses VP1 protein |
| title | Molecular strategy for the direct detection and identification of human enteroviruses in clinical specimens associated with hand, foot and mouth disease |
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