{"id":13840,"date":"2026-05-13T14:43:17","date_gmt":"2026-05-13T06:43:17","guid":{"rendered":"https:\/\/vibrojet-tech.com\/?post_type=docs&p=13840"},"modified":"2026-05-25T15:04:37","modified_gmt":"2026-05-25T07:04:37","password":"","slug":"3%ef%bd%885%e4%ba%9a%e5%9e%8b%e9%ab%98%e8%87%b4%e7%97%85%e6%80%a7%e7%a6%bd%e6%b5%81%e6%84%9f%e7%97%85%e6%af%92%e5%be%ae%e6%bb%b4%e5%bc%8f%e6%95%b0%e5%ad%97pcr%e6%a3%80%e6%b5%8b%e6%96%b9%e6%b3%95","status":"publish","type":"docs","link":"https:\/\/vibrojet-tech.com\/?docs=3%ef%bd%885%e4%ba%9a%e5%9e%8b%e9%ab%98%e8%87%b4%e7%97%85%e6%80%a7%e7%a6%bd%e6%b5%81%e6%84%9f%e7%97%85%e6%af%92%e5%be%ae%e6%bb%b4%e5%bc%8f%e6%95%b0%e5%ad%97pcr%e6%a3%80%e6%b5%8b%e6%96%b9%e6%b3%95","title":{"rendered":"\uff285 \u4e9a\u578b\u9ad8\u81f4\u75c5\u6027\u79bd\u6d41\u611f\u75c5\u6bd2\u5fae\u6ef4\u5f0f\u6570\u5b57 PCR \u68c0\u6d4b\u65b9\u6cd5\u7684\u5efa\u7acb\u4e0e\u5e94\u7528"},"content":{"rendered":"\n
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<\/a><\/a>\u300a\u4e2d\u56fd\u517d\u533b\u5b66\u62a5\u300b<\/p>\n\n\n\n

2023\u5e74\u7b2c5\u671f<\/p>\n<\/div>\n\n\n\n

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DOI:<\/p>\n\n\n\n

<\/a><\/a>10.16303\/j.cnki.1005-4545.2023.05.16<\/p>\n<\/div>\n\n\n\n

\u848b\u6587\u660e\uff0c\u5218\u6714\uff0c\u674e\u91d1\u5e73\uff0c\u5f6d\u7a0b\uff0c\u5c39\u99a8\uff0c\u4e8e\u6653\u6167\uff0c\u674e\u9633\uff0c\u738b\u9759\u9759\uff0c\u5218\u534e\u96f7<\/p>\n\n\n\n

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Abstract<\/h2>\n\n\n\n

\u4e3a\u5efa\u7acb\u7279\u5f02\u3001\u7075\u654f\u7684 H5 \u4e9a\u578b\u9ad8\u81f4\u75c5\u6027\u79bd\u6d41\u611f\u75c5\u6bd2\uff08high pathogenicity avian influenza virus, HPAIV\uff09\u5fae\u6ef4\u5f0f\u6570\u5b57 PCR\uff08droplet digital PCR, ddPCR\uff09\u68c0\u6d4b\u65b9\u6cd5\uff0c\u6839\u636e GISAID \u548c GenBank \u516c\u5e03\u7684 H5 \u4e9a\u578b HPAIVs HA<\/em> \u57fa\u56e0\u7684\u6838\u82f7\u9178\u5e8f\u5217\uff0c\u8bbe\u8ba1\u5e76\u5408\u6210\u5f15\u7269\u548c\u63a2\u9488\uff0c\u5bf9\u53cd\u5e94\u4f53\u7cfb\u548c\u6761\u4ef6\u8fdb\u884c\u4f18\u5316\uff0c\u5efa\u7acb\u9488\u5bf9 H5 \u4e9a\u578b HPAIVs \u7684 ddPCR \u68c0\u6d4b\u65b9\u6cd5\uff0c\u5e76\u5bf9\u8be5\u65b9\u6cd5\u7684\u7279\u5f02\u6027\u3001\u91cd\u590d\u6027\u4ee5\u53ca\u654f\u611f\u6027\u8fdb\u884c\u8bc4\u4f30\u3002\u7ed3\u679c\u663e\u793a\uff0c\u5f15\u7269\u63a2\u9488\u6d53\u5ea6\u4e3a900 nmol\/L\u548c250 nmol\/L\u3001\u9000\u706b\u6e29\u5ea6\u4e3a55 \u2103\u3001\u5347\u964d\u6e29\u901f\u7387\u4e3a2.0 \u2103\/s\u65f6 ddPCR \u53cd\u5e94\u6548\u7387\u6700\u9ad8\u3002\u8be5\u65b9\u6cd5\u53ef\u7279\u5f02\u6027\u68c0\u6d4b H5 \u4e9a\u578b HPAIVs\uff0c\u5bf9 H9N2\u3001H7N9 \u7b49\u5e38\u89c1\u79bd\u75c5\u75c5\u539f\u5747\u4e0d\u80fd\u68c0\u51fa\u3002\u8be5\u65b9\u6cd5\u5bf9\u6807\u51c6\u54c1\u6700\u4f4e\u68c0\u6d4b\u9650\u4e3a1.97 \u62f7\u8d1d \/\u03bcL\uff0c\u654f\u611f\u6027\u6bd4 qPCR \u65b9\u6cd5\u9ad8 10 \u500d\u3002\u7ec4\u5185\u548c\u7ec4\u95f4\u91cd\u590d\u6027\u8bd5\u9a8c\u7684\u53d8\u5f02\u7cfb\u6570\u5747\u5c0f\u4e8e5 %\u3002\u5bf9 80 \u4efd\u4e34\u5e8a\u6837\u54c1\u68c0\u6d4b\u7ed3\u679c\u663e\u793a\uff0c\u4e0e\u91d1\u6807\u51c6\u65b9\u6cd5\u7684\u7b26\u5408\u7387\u4e3a100 %\u3002\u672c\u7814\u7a76\u5efa\u7acb\u7684 ddPCR \u65b9\u6cd5\u7279\u5f02\u6027\u5f3a\u3001\u654f\u611f\u6027\u9ad8\u3001\u91cd\u590d\u6027\u597d\uff0c\u4e3a H5 \u4e9a\u578b HPAIVs \u7684\u8bca\u65ad\u548c\u68c0\u6d4b\u4ee5\u53ca\u7cbe\u51c6\u9632\u63a7\u63d0\u4f9b\u4e86\u91cd\u8981\u6280\u672f\u652f\u6491\u3002<\/p>\n\n\n\n

To establish a specific and sensitive droplet digital PCR (ddPCR) method for detection of H5 subtype high pathogenicity avian influenza virus (HPAIV), primers and probes were designed and synthesized based on the nucleotide sequences of H5 subtype HPAIV HA<\/em> genes published in GISAID and GenBank. The reaction system and conditions were optimized, and the specificity, repeatability and sensitivity of the method were systematically evaluated. The results showed that the optimal concentrations of primers and probe were 900 nmol\/L and 250 nmol\/L, respectively. The optimal annealing temperature was 55 \u2103, and the optimal heating and cooling rate was 2.0 \u2103\/s. The method could specifically detect H5 subtype HPAIV, but showed no cross-reactivity with H9N2 and H7N9 subtype AIVs or other common avian pathogens. The limit of detection for the standard substance was 1.97 copies\/\u03bcL, and the sensitivity of this method was 10 times higher than that of qPCR. The coefficient of variation for both intra- and inter-assay repeatability tests was less than 5 %. The detection results of 80 clinical samples showed that the coincidence rate with the gold standard method was 100 %. The ddPCR method established in this study has strong specificity, high sensitivity and good repeatability, which provides important technical support for the diagnosis, detection and precise prevention and control of H5 subtype HPAIVs.<\/p>\n\n\n\n

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Tag<\/strong>s:<\/h2>\n\n\n\n

\u79bd\u6d41\u611f\u75c5\u6bd2\uff08avian influenza virus\uff09\uff1bH5\u4e9a\u578b\u9ad8\u81f4\u75c5\u6027\u79bd\u6d41\u611f\u75c5\u6bd2\uff08H5 subtype high pathogenicity avian influenza virus\uff09\uff1b\u5fae\u6ef4\u5f0f\u6570\u5b57PCR\uff08droplet digit al PCR\uff09<\/p>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"

Establishes a ddPCR assay for highly pathogenic H5 avian influenza virus, supporting digital PCR use in animal disease pathogen detection.<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_eb_attr":"","site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"disabled","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"advanced_seo_description":"","jetpack_seo_html_title":"","jetpack_seo_noindex":false,"footnotes":""},"doc_category":[30],"doc_tag":[],"class_list":["post-13840","docs","type-docs","status-publish","hentry","doc_category-platform-analytical-performance"],"year_month":"2026-05","word_count":268,"total_views":0,"reactions":{"happy":0,"normal":0,"sad":0},"author_info":{"name":"root","author_nicename":"root","author_url":"https:\/\/vibrojet-tech.com\/?author=1"},"doc_category_info":[{"term_name":"Platform Analytical Performance","term_url":"https:\/\/vibrojet-tech.com\/?doc_category=platform-analytical-performance"}],"doc_tag_info":[],"jetpack_sharing_enabled":true,"_links":{"self":[{"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/docs\/13840","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/docs"}],"about":[{"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/types\/docs"}],"author":[{"embeddable":true,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=13840"}],"version-history":[{"count":19,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/docs\/13840\/revisions"}],"predecessor-version":[{"id":14755,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/docs\/13840\/revisions\/14755"}],"wp:attachment":[{"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=13840"}],"wp:term":[{"taxonomy":"doc_category","embeddable":true,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=%2Fwp%2Fv2%2Fdoc_category&post=13840"},{"taxonomy":"doc_tag","embeddable":true,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=%2Fwp%2Fv2%2Fdoc_tag&post=13840"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}