Jie CHE , Bo Han CHEN , Li XU , Yuan GAO , Meng Meng YUE , Zi Man CHEN , Mao Jun ZHANG , Zhu Jun SHAO
{"title":"基于定量实时聚合酶链反应的97种肺炎球菌血清学分型方法的建立和改进。","authors":"Jie CHE , Bo Han CHEN , Li XU , Yuan GAO , Meng Meng YUE , Zi Man CHEN , Mao Jun ZHANG , Zhu Jun SHAO","doi":"10.3967/bes2023.078","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes.</p></div><div><h3>Methods</h3><p>A database of capsular polysaccharide (<em>cps</em>) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the <em>cps</em> loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping.</p></div><div><h3>Results</h3><p>A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1–100 copies/μL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains.</p></div><div><h3>Conclusion</h3><p>A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.</p></div>","PeriodicalId":55364,"journal":{"name":"Biomedical and Environmental Sciences","volume":"36 9","pages":"Pages 787-799"},"PeriodicalIF":3.0000,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Establishment and Modification of Ninety-seven Pneumococcal Serotyping Assays Based on Quantitative Real-time Polymerase Chain Reaction\",\"authors\":\"Jie CHE , Bo Han CHEN , Li XU , Yuan GAO , Meng Meng YUE , Zi Man CHEN , Mao Jun ZHANG , Zhu Jun SHAO\",\"doi\":\"10.3967/bes2023.078\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><p>To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes.</p></div><div><h3>Methods</h3><p>A database of capsular polysaccharide (<em>cps</em>) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the <em>cps</em> loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping.</p></div><div><h3>Results</h3><p>A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1–100 copies/μL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains.</p></div><div><h3>Conclusion</h3><p>A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.</p></div>\",\"PeriodicalId\":55364,\"journal\":{\"name\":\"Biomedical and Environmental Sciences\",\"volume\":\"36 9\",\"pages\":\"Pages 787-799\"},\"PeriodicalIF\":3.0000,\"publicationDate\":\"2023-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biomedical and Environmental Sciences\",\"FirstCategoryId\":\"1089\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0895398823001319\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"ENVIRONMENTAL SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biomedical and Environmental Sciences","FirstCategoryId":"1089","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0895398823001319","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ENVIRONMENTAL SCIENCES","Score":null,"Total":0}
Establishment and Modification of Ninety-seven Pneumococcal Serotyping Assays Based on Quantitative Real-time Polymerase Chain Reaction
Objective
To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes.
Methods
A database of capsular polysaccharide (cps) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the cps loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping.
Results
A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1–100 copies/μL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains.
Conclusion
A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.
期刊介绍：
Biomedical and Environmental Sciences (BES) is a peer-reviewed journal jointly established by the Chinese Center for Disease Control and Prevention (China CDC) and the Coulston International Corporation (CIC), USA in 1988, and is published monthly by Elsevier. It is indexed by SCI, PubMed, and CA.
Topics covered by BES include infectious disease prevention, chronic and non-communicable disease prevention, disease control based on preventive medicine, and public health theories. It also focuses on the health impacts of environmental factors in people''s daily lives and work, including air quality, occupational hazards, and radiation hazards.
Article types considered for publication include original articles, letters to the editor, reviews, research highlights, and policy forum.
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