{"id":13842,"date":"2026-05-13T14:45:35","date_gmt":"2026-05-13T06:45:35","guid":{"rendered":"https:\/\/vibrojet-tech.com\/?post_type=docs&p=13842"},"modified":"2026-05-25T15:07:03","modified_gmt":"2026-05-25T07:07:03","password":"","slug":"4%ef%bd%887%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=4%ef%bd%887%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":"\uff287 \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>\u300a\u4e2d\u56fd\u517d\u533b\u6742\u5fd7\u300b<\/p>\n\n\n\n

2023\u5e74\u7b2c2\u671f<\/p>\n<\/div>\n\n\n\n

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

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

\u848b\u6587\u660e\uff0c\u674e\u91d1\u5e73\uff0c\u5c39\u99a8\uff0c\u5f6d\u7a0b\uff0c\u5218\u6714\uff0c\u674e\u9633\uff0c\u4e8e\u6653\u6167\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\u7075\u654f\u3001\u7279\u5f02\u7684 H7 \u4e9a\u578b\u79bd\u6d41\u611f\u75c5\u6bd2\uff08AIV\uff09\u5fae\u6ef4\u5f0f\u6570\u5b57 PCR\uff08ddPCR\uff09\u68c0\u6d4b\u65b9\u6cd5\uff0c\u672c\u7814\u7a76\u4f9d\u636e GenBank \u548c GISAID \u6536\u5f55\u7684 H7 \u4e9a\u578b AIV \u8840\u51dd\u7d20\uff08HA<\/em>\uff09\u57fa\u56e0\u6838\u82f7\u9178\u5e8f\u5217\uff0c\u8bbe\u8ba1\u5e76\u5408\u6210 H7 \u4e9a\u578b AIV \u7279\u5f02\u6027\u5f15\u7269\u4e0e\u63a2\u9488\uff0c\u4f18\u5316\u6269\u589e\u53cd\u5e94\u6761\u4ef6\uff0c\u5efa\u7acb H7 \u4e9a\u578b AIV ddPCR \u68c0\u6d4b\u4f53\u7cfb\uff0c\u5e76\u8bc4\u4ef7\u5176\u7279\u5f02\u6027\u3001\u654f\u611f\u6027\u4e0e\u91cd\u590d\u6027\u3002\u7ed3\u679c\u663e\u793a\uff1a\u6700\u4f73\u5f15\u7269\u6d53\u5ea6\u4e3a900 nmol\/L\u3001\u63a2\u9488\u6d53\u5ea6\u4e3a250 nmol\/L\uff0c\u6700\u4f73\u9000\u706b\u6e29\u5ea655 \u2103\uff0c\u6700\u4f73\u5347\u964d\u6e29\u901f\u73872.0 \u2103\/s\uff0c\u6700\u4f73\u53cd\u8f6c\u5f55\u65f6\u95f420 min\u3002\u7279\u5f02\u6027\u8bd5\u9a8c\u8868\u660e\uff0c\u8be5\u65b9\u6cd5\u4ec5\u53ef\u7279\u5f02\u6027\u68c0\u51fa H7 \u4e9a\u578b AIV\uff0c\u65e0\u6cd5\u68c0\u51fa H5 \u4e9a\u578b AIV\u3001H9 \u4e9a\u578b AIV\u3001\u65b0\u57ce\u75ab\u75c5\u6bd2\u3001\u9e21\u4f20\u67d3\u6027\u652f\u6c14\u7ba1\u708e\u75c5\u6bd2\u53ca\u4f20\u67d3\u6027\u6cd5\u6c0f\u56ca\u75c5\u6bd2\u3002\u654f\u611f\u6027\u8bd5\u9a8c\u7ed3\u679c\u663e\u793a\uff0c\u8be5\u65b9\u6cd5\u5bf9\u91cd\u7ec4\u8d28\u7c92\u6807\u51c6\u54c1\u7684\u6700\u4f4e\u68c0\u6d4b\u4e0b\u9650\u4e3a0.8 copies\/\u03bcL\uff0c\u7075\u654f\u5ea6\u8f83\u8367\u5149\u5b9a\u91cf PCR\uff08qPCR\uff09\u9ad8\u51fa 10 \u500d\u3002\u7ec4\u5185\u4e0e\u7ec4\u95f4\u91cd\u590d\u6027\u8bd5\u9a8c\u53d8\u5f02\u7cfb\u6570\u5747\u5c0f\u4e8e3 %\u3002\u5bf9 60 \u4efd\u4e34\u5e8a\u6837\u54c1\u5f00\u5c55\u68c0\u6d4b\uff0c\u8be5\u65b9\u6cd5\u4e0e\u9e21\u80da\u5206\u79bb\u6cd5\u68c0\u6d4b\u7b26\u5408\u7387\u8fbe100 %\u3002\u7efc\u4e0a\uff0c\u672c\u7814\u7a76\u5efa\u7acb\u7684 H7 \u4e9a\u578b AIV ddPCR \u68c0\u6d4b\u65b9\u6cd5\u7279\u5f02\u6027\u5f3a\u3001\u91cd\u590d\u6027\u597d\u3001\u7075\u654f\u5ea6\u9ad8\uff0c\u53ef\u5e94\u7528\u4e8e H7 \u4e9a\u578b AIV \u65e9\u671f\u8bca\u65ad\u3001\u6d41\u884c\u75c5\u5b66\u8c03\u67e5\u3001\u68c0\u6d4b\u65b9\u6cd5\u8bc4\u4ef7\u3001\u6838\u9178\u6807\u51c6\u7269\u8d28\u7814\u5236\u53ca\u514d\u75ab\u6548\u679c\u8bc4\u4f30\uff0c\u53ef\u4e3a H7 \u4e9a\u578b AIV \u79d1\u5b66\u9632\u63a7\u4e0e\u76f8\u5173\u57fa\u7840\u7814\u7a76\u63d0\u4f9b\u53ef\u9760\u6280\u672f\u652f\u6491\u3002<\/p>\n\n\n\n

In order to establish a sensitive and specific droplet digital PCR (ddPCR) method for the detection of H7 subtype avian influenza virus (AIV), this study designed and synthesized primers and probe according to the nucleotide sequence of H7 subtype AIV HA<\/em> genes in GenBank and GISAID, optimized the reaction conditions to establish a ddPCR method, and determined the specificity, sensitivity and repeatability. The results showed that the optimal primers and probe concentration were 900 nmol\/L and 250 nmol\/L, respectively; the optimal annealing temperature was 55 \u2103; the optimal rising and cooling rate was 2.0 \u2103\/s; and the optimal time of reverse transcription was 20 min. The specificity test results showed that this method detected only H7 subtype AIV, but not H5 subtype AIV, H9 subtype AIV, Newcastle disease virus, infectious bronchitis virus and infectious bursal disease virus. Sensitivity test results showed that the detection limit of this method was 0.8 copies\/\u03bcL, which was 10 times more sensitive than qPCR. The coefficients of variation were less than 3 % in both intra-batch and inter-batch reproducibility tests. The results of 60 clinical samples showed that the coincidence rate was 100 % compared with the virus isolation method. Thus, the ddPCR method established in this study has excellent specificity, high sensitivity and good reproducibility, allows for the early diagnosis, epidemiological investigation, diagnostic method evaluation, standard substance development and immune effect evaluation of H7 subtype AIV, and provides strong technical support for the prevention and control of H7 subtype AIV.<\/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\uff1bH7 \u4e9a\u578b\u9ad8\u81f4\u75c5\u6027\u79bd\u6d41\u611f\u75c5\u6bd2\uff08H7 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 H7 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-13842","docs","type-docs","status-publish","hentry","doc_category-platform-analytical-performance"],"year_month":"2026-05","word_count":295,"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\/13842","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=13842"}],"version-history":[{"count":21,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/docs\/13842\/revisions"}],"predecessor-version":[{"id":14757,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=\/wp\/v2\/docs\/13842\/revisions\/14757"}],"wp:attachment":[{"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=13842"}],"wp:term":[{"taxonomy":"doc_category","embeddable":true,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=%2Fwp%2Fv2%2Fdoc_category&post=13842"},{"taxonomy":"doc_tag","embeddable":true,"href":"https:\/\/vibrojet-tech.com\/index.php?rest_route=%2Fwp%2Fv2%2Fdoc_tag&post=13842"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}