{"id":13884,"date":"2026-05-13T17:16:16","date_gmt":"2026-05-13T09:16:16","guid":{"rendered":"https:\/\/vibrojet-tech.com\/?post_type=docs&p=13884"},"modified":"2026-05-25T15:57:42","modified_gmt":"2026-05-25T07:57:42","password":"","slug":"%e5%86%85%e5%90%ab%e8%b5%a4%e7%be%bd%e7%97%85%e7%97%85%e6%af%92s%e5%9f%ba%e5%9b%a0%e5%81%87%e7%97%85%e6%af%92%e7%9a%84%e6%9e%84%e5%bb%ba%e3%80%81%e9%89%b4%e5%ae%9a%e5%8f%8a%e5%88%9d%e6%ad%a5%e5%ba%94","status":"publish","type":"docs","link":"https:\/\/vibrojet-tech.com\/?docs=%e5%86%85%e5%90%ab%e8%b5%a4%e7%be%bd%e7%97%85%e7%97%85%e6%af%92s%e5%9f%ba%e5%9b%a0%e5%81%87%e7%97%85%e6%af%92%e7%9a%84%e6%9e%84%e5%bb%ba%e3%80%81%e9%89%b4%e5%ae%9a%e5%8f%8a%e5%88%9d%e6%ad%a5%e5%ba%94","title":{"rendered":"\u5185\u542b\u8d64\u7fbd\u75c5\u75c5\u6bd2 S \u57fa\u56e0\u5047\u75c5\u6bd2\u7684\u6784\u5efa\u3001\u9274\u5b9a\u53ca\u521d\u6b65\u5e94\u7528"},"content":{"rendered":"\n
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<\/a>\u300a\u4e2d\u56fd\u52a8\u7269\u68c0\u75ab\u300b<\/p>\n\n\n\n

2025\u5e74\u7b2c1\u671f<\/p>\n<\/div>\n\n\n\n

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

<\/a><\/a>10.3969\/j.issn.1005-944X.2025.01.016<\/p>\n<\/div>\n\n\n\n

\u674e\u8d85\uff1b\u6f58\u4fca\u6167\uff1b\u738b\u7d20\u6625\uff1b\u968b\u91d1\u94b0\uff1b\u9b4f\u4e16\u840c\uff1b\u7941\u5029\uff1b\u5434\u53d1\u5174\uff1b\u674e\u535a\u6587\uff1b\u738b\u6977\u5bac<\/p>\n\n\n\n

\n\n\n\n

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

\u4e3a\u5236\u5907\u8d64\u7fbd\u75c5\u75c5\u6bd2\uff08Akabane virus, AKAV\uff09\u6838\u9178\u8bca\u65ad\u8bd5\u5242\u9633\u6027\u5bf9\u7167\u54c1\uff0c\u672c\u7814\u7a76\u5148\u5408\u6210 AKAV S \u5168\u957f\u57fa\u56e0\uff0c\u6784\u5efa\u91cd\u7ec4\u6162\u75c5\u6bd2\u7a7f\u68ad\u8f7d\u4f53 pLenti-GIII-CMV-CBH-GFP-2A-Puro-AKAV\u3002\u5c06\u6d4b\u5e8f\u5408\u683c\u7684\u91cd\u7ec4\u8f7d\u4f53\u4e0e\u5305\u88c5\u8f7d\u4f53\u5171\u8f6c\u67d3 293T \u7ec6\u80de\uff0c\u6536\u96c6\u5e76\u7eaf\u5316\u7ec6\u80de\u57f9\u517b\u6db2\u4e0a\u6e05\uff0c\u68c0\u6d4b\u8d28\u7c92\u6838\u9178\u6b8b\u7559\u3002\u5229\u7528\u4e0a\u6e05\u4fb5\u67d3 293T \u7ec6\u80de\uff0c\u89c2\u6d4b\u7eff\u8272\u8367\u5149\u86cb\u767d\u8868\u8fbe\uff0c\u901a\u8fc7 RT-PCR\u3001\u8367\u5149\u5b9a\u91cf RT-PCR \u9274\u5b9a AKAV S \u57fa\u56e0\u7247\u6bb5\uff0c\u6700\u7ec8\u91c7\u7528\u6570\u5b57 RT-PCR \u5b8c\u6210\u7edd\u5bf9\u5b9a\u91cf\u3002\u7ed3\u679c\u663e\u793a\uff0c\u7eaf\u5316\u4e0a\u6e05\u672a\u68c0\u51fa\u8d28\u7c92\u6b8b\u7559\uff1b\u57f9\u517b72 h\u7684 293T \u7ec6\u80de\u53ef\u89c1\u7eff\u8272\u8367\u5149\uff1b\u4e24\u79cd PCR \u65b9\u6cd5\u5747\u53ef\u6269\u589e\u51fa\u76ee\u7684\u57fa\u56e0\u7247\u6bb5\uff1b\u5047\u75c5\u6bd2\u62f7\u8d1d\u6570\u6d53\u5ea6\u5b9a\u503c\u4e3a3.36 \u00d7 10\u00b3 copies\/\u03bcL\u3002\u672c\u7814\u7a76\u6210\u529f\u6784\u5efa\u643a\u5e26 AKAV S \u5168\u957f\u57fa\u56e0\u7684\u5047\u75c5\u6bd2\uff0c\u53ef\u4f5c\u4e3a\u6838\u9178\u68c0\u6d4b\u9633\u6027\u5bf9\u7167\u54c1\uff0c\u4e3a\u8d64\u7fbd\u75c5\u75c5\u6bd2\u6838\u9178\u8bca\u65ad\u8bd5\u5242\u76d2\u7814\u53d1\u63d0\u4f9b\u652f\u6491\u3002<\/p>\n\n\n\n

In order to prepare positive control materials for Akabane virus (AKAV) nucleic acid diagnostic reagents, the full-length AKAV S gene was synthesized. The verified recombinant lentiviral shuttle vector was co-transfected into 293T cells together with packaging vectors. Purified cell culture supernatant was collected and tested for plasmid residue. The supernatant was used to infect 293T cells, and green fluorescent protein expression was observed. RT-PCR and quantitative real-time RT-PCR were applied to identify AKAV S gene fragments, and digital RT-PCR was adopted for absolute quantification. No plasmid residue was detected in purified supernatant. Green fluorescence was observed in 293T cells cultured for 72 hours. Target AKAV S gene fragments were successfully amplified by both PCR methods. The quantified concentration of pseudovirus solution was 3.36 \u00d7 10\u00b3 copies\/\u03bcL. The pseudovirus carrying full-length AKAV S gene was successfully constructed. It can serve as positive control for AKAV nucleic acid detection and lays a foundation for relevant diagnostic kit development.<\/p>\n\n\n\n

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

\u8d64\u7fbd\u75c5\u75c5\u6bd2\uff08AKAV\uff09\uff1bS\u57fa\u56e0\uff08S gene\uff09\uff1b \u5047\u75c5\u6bd2\u7c92\u5b50\uff08pseudovirion\uff09\uff1b \u9633\u6027\u5bf9\u7167\u54c1\uff08positive control material\uff09<\/p>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"

Constructs and evaluates a pseudovirus containing the Akabane virus S gene, supporting quality-control material development and method validation for animal viral nucleic acid 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