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Article(id=1307007459927806236, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, articleNumber=null, orderNo=null, doi=10.11843/j.issn.0366-6964.2026.01.049, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1733932800000, receivedDateStr=2024-12-12, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1789544461470, onlineDateStr=2026-09-16, pubDate=1769097600000, pubDateStr=2026-01-23, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789544461470, onlineIssueDateStr=2026-09-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789544461470, creator=13701087609, updateTime=1789544461470, updator=13701087609, issue=Issue{id=1307007396681900041, tenantId=1146029695717560320, journalId=1306274286668972124, year='2026', volume='57', issue='1', pageStart='1', pageEnd='568', issueExtLink='null', onlineDate='null', pubDate='1769097600000', pubDateStr='2026-01-23', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1789544446392, creator='13701087609', updateTime=1789609890784, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1307281890638057732, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1307281890738721029, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=564, endPage=568, ext={EN=ArticleExt(id=1307007462817681696, articleId=1307007459927806236, tenantId=1146029695717560320, journalId=1306274286668972124, language=EN, title=Development and Evaluation of the Multienzyme Isothermal Rapid Amplification Assays for Detection of Streptococcus suis and Its Three Serovars, columnId=1307007462452777247, journalTitle=Acta Veterinaria et Zootechnica Sinica, columnName=RESEARCH NOTES, runingTitle=null, highlight=null, articleAbstract=

Streptococcus suis （SS） is one of the important pathogens causing serious economic losses in the pig farming industry. Here， we developed a novel rapid detection method for detecting SS and serovars 2， 7 and 9 based on multienzyme isothermal rapid amplification （MIRA） technology. The MIRA assays were validated for detection of SS and it serovars within 5 to 30 min at 20-41 ℃， with relative higher specificity and sensitivity. There are 100% agreement between our MIRA and PCR methods in the detection of clinical samples. Our results showed that the MIRA was a useful diagnostic method for rapid diagnosis of SS infection.

, authors=Zhangyu HU, Yongqing ZHANG, Yilian CHEN, Fazhi XU, Xinyue CHANG, Fugui FANG, Liang LI, Pei SUN, Xuelan LIU, authorsList=Zhangyu HU, Yongqing ZHANG, Yilian CHEN, Fazhi XU, Xinyue CHANG, Fugui FANG, Liang LI, Pei SUN, Xuelan LIU, authorCompany=null, correspAuthors=Xuelan LIU, authorNote=null, correspAuthorsNote=
LIU Xuelan， E-mail：
, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1307007472015790380, articleId=1307007459927806236, tenantId=1146029695717560320, journalId=1306274286668972124, language=CN, title=基于多酶恒温快速扩增检测猪链球菌及其3种主要血清型方法的建立及应用, columnId=1307007463681708323, journalTitle=畜牧兽医学报, columnName=研究简报, runingTitle=null, highlight=null, articleAbstract=

猪链球菌（Streptococcus suis，SS）影响养猪业的重要病原菌之一。本文基于多酶恒温快速扩增（multienzyme isothermal rapid amplification，MIRA）技术建立快速检测SS及其血清2、7、9型3种主要临床血清型的新方法，该方法可在20~41 ℃温度下5~30 min内完成扩增，具有良好的特异性和敏感性；应用MIRA方法能够快速检测临床分离菌株中相应血清型，且与PCR方法的符合率为100%。上述结果表明，本研究所建立的MIRA方法可为临床诊断猪链球菌感染提供简便快捷的方法。

, authors=胡章玉, 张永青, 陈忆廉, 许发芝, 常馨月, 方富贵, 李亮, 孙裴, 刘雪兰, authorsList=胡章玉, 张永青, 陈忆廉, 许发芝, 常馨月, 方富贵, 李亮, 孙裴, 刘雪兰, authorCompany=null, correspAuthors=刘雪兰, authorNote=

胡章玉和张永青是同等贡献作者

胡章玉，本科生，主要从事病原微生物学研究，E-mail：

张永青，硕士生，主要从事病原微生物学研究，E-mail：

, correspAuthorsNote=
刘雪兰，主要从事病原微生物与免疫学研究，E-mail：
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胡章玉，本科生，主要从事病原微生物学研究，E-mail：

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胡章玉，本科生，主要从事病原微生物学研究，E-mail：

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张永青，硕士生，主要从事病原微生物学研究，E-mail：

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M. DL2000 DNA marker； A-D.MIRA primer screening for SS （A）， SS2 （B）， SS7（C） or SS9（D）， respectively； 1-4. Products of MIRA amplification using different primers. NC. ddH2O as template for MIRA control

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M.DL2000 DNA相对分子质量标准；A~D.分别为SS、SS2、SS7与SS9血清型的MIRA扩增结果；1~4.利用不同引物进行MIRA扩增的结果；NC.ddH2O为模板的MIRA对照

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Primers for MIRA detection of SS and 3 serovars

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SS及其血清型

SS and serovars

名称

Name of primer

引物序列（5′→3′）

Primer sequence

片段长度/bp

Length

SSSS-F1/R1

CAGAAACTGTCTTTATGCGATTGATT

AGTCATTAAATTGTGGACGAGAGAGCT

427
SS-F2/R2

GTAGGCTAACTAAAGCGGGCGATTGG

ATTACGGGGTCATTTTGCCGAGTTCC

262
SS-F3/R3

CCCATCCACCGTAAGACTAAGGTTTC

CTTAGTTAGCCTACTGCGTCCCTCCA

156
SS2SS2-F1/R1

GGGGTTCGTATTAACTTACTTGGCGTA

TTAAGGGCATTTCAAGCGTTCCATTC

433
SS2-F2/R2

ACTTTAGACTATGCTATAAGGGTTAGACGC

TTCTACTGTTGCAGATAGGACTACAATGAT

222
SS2-F3/R3

TTTAGGTTGGAATGGAACGCTTGAAATG

CTCTACTGTTGCAGATAGGACTACAATG

188
SS7SS7-F1/R1

TAGAGGTAGGTGGATTGCGTTCTTG

AACACCCTGGATAGCCGTAATTGTA

215
SS7-F2/R2

TAATAATGCCCTCGTGGAATACAGCTAAGT

GGTGCCATAAATCATCTGAATCCAAGAACG

267
SS7-F3/R3

TAGAGGTAGGTGGATTGCGTTCTTGG

ATTCAAACACCCTGGATAGCCGTAATT

218
SS7-F4/R4

TTAGTAGTAGGTGCTGGAGATGGTGGG

ATGTTACCCGCCATAATGTCTTCAATGC

310
SS9SS9-F1/R1

TATCAGACTGCATACAGGGCAGAAG

ATCACAAGCAACCTGGGCAACAAAA

282
SS9-F2/R2

TTGTCTCTCAAAATAAATCTAATCAGGGGC

GAATTGTGAAACGACTTGCTACTAGAGGAT

490
SS9-F3/R3

GGAAGTATTGGCAACGAAGAAGGGA

CACATCATCAACAGCCTCTATATCCGC

177
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用于MIRA检测SS及3种血清型的引物

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SS及其血清型

SS and serovars

名称

Name of primer

引物序列（5′→3′）

Primer sequence

片段长度/bp

Length

SSSS-F1/R1

CAGAAACTGTCTTTATGCGATTGATT

AGTCATTAAATTGTGGACGAGAGAGCT

427
SS-F2/R2

GTAGGCTAACTAAAGCGGGCGATTGG

ATTACGGGGTCATTTTGCCGAGTTCC

262
SS-F3/R3

CCCATCCACCGTAAGACTAAGGTTTC

CTTAGTTAGCCTACTGCGTCCCTCCA

156
SS2SS2-F1/R1

GGGGTTCGTATTAACTTACTTGGCGTA

TTAAGGGCATTTCAAGCGTTCCATTC

433
SS2-F2/R2

ACTTTAGACTATGCTATAAGGGTTAGACGC

TTCTACTGTTGCAGATAGGACTACAATGAT

222
SS2-F3/R3

TTTAGGTTGGAATGGAACGCTTGAAATG

CTCTACTGTTGCAGATAGGACTACAATG

188
SS7SS7-F1/R1

TAGAGGTAGGTGGATTGCGTTCTTG

AACACCCTGGATAGCCGTAATTGTA

215
SS7-F2/R2

TAATAATGCCCTCGTGGAATACAGCTAAGT

GGTGCCATAAATCATCTGAATCCAAGAACG

267
SS7-F3/R3

TAGAGGTAGGTGGATTGCGTTCTTGG

ATTCAAACACCCTGGATAGCCGTAATT

218
SS7-F4/R4

TTAGTAGTAGGTGCTGGAGATGGTGGG

ATGTTACCCGCCATAATGTCTTCAATGC

310
SS9SS9-F1/R1

TATCAGACTGCATACAGGGCAGAAG

ATCACAAGCAACCTGGGCAACAAAA

282
SS9-F2/R2

TTGTCTCTCAAAATAAATCTAATCAGGGGC

GAATTGTGAAACGACTTGCTACTAGAGGAT

490
SS9-F3/R3

GGAAGTATTGGCAACGAAGAAGGGA

CACATCATCAACAGCCTCTATATCCGC

177
), ArticleFig(id=1307286552615347047, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007459927806236, language=EN, label=Table 2, caption=

Detection of SS Clinical isolates using MIRA or PCR assays

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检测方法

Detection method

总数（n）TotalSS阳性数（n）SS positiveSS2阳性数（n）SS2 positiveSS7阳性数（n）SS7 positiveSS9阳性数（n）SS9 positive

符合率/%

Coincidence rate

MIRA36302110100
PCR36302110100
), ArticleFig(id=1307286552686650216, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007459927806236, language=CN, label=表2, caption=

应用MIRA检测SS临床分离菌株的结果

, figureFileSmall=null, figureFileBig=null, tableContent=

检测方法

Detection method

总数（n）TotalSS阳性数（n）SS positiveSS2阳性数（n）SS2 positiveSS7阳性数（n）SS7 positiveSS9阳性数（n）SS9 positive

符合率/%

Coincidence rate

MIRA36302110100
PCR36302110100
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胡章玉 , 张永青 , 陈忆廉 , 许发芝 , 常馨月 , 方富贵 , 李亮 , 孙裴 , 刘雪兰
畜牧兽医学报 | 研究简报 2026,57(1): 564-568
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畜牧兽医学报 |研究简报 2026 , 57 (1) : 564 -568
基于多酶恒温快速扩增检测猪链球菌及其3种主要血清型方法的建立及应用
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胡章玉，本科生，主要从事病原微生物学研究，E-mail：

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胡章玉 , 张永青 , 陈忆廉, 许发芝, 常馨月, 方富贵, 李亮, 孙裴, 刘雪兰
作者信息
  • 安徽农业大学动物医学院，合肥 230036
通讯作者:
刘雪兰，主要从事病原微生物与免疫学研究，E-mail：
作者简介:

胡章玉和张永青是同等贡献作者

胡章玉，本科生，主要从事病原微生物学研究，E-mail：

张永青，硕士生，主要从事病原微生物学研究，E-mail：

Development and Evaluation of the Multienzyme Isothermal Rapid Amplification Assays for Detection of Streptococcus suis and Its Three Serovars
Zhangyu HU , Yongqing ZHANG , Yilian CHEN, Fazhi XU, Xinyue CHANG, Fugui FANG, Liang LI, Pei SUN, Xuelan LIU
Affiliations
  • College of Veterinary Medicine，Anhui Agricultural University，Hefei 230036，China
    • Corresponding Author:
    LIU Xuelan， E-mail：
出版时间: 2026-01-23 doi: 10.11843/j.issn.0366-6964.2026.01.049
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摘要
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猪链球菌（Streptococcus suis，SS）影响养猪业的重要病原菌之一。本文基于多酶恒温快速扩增（multienzyme isothermal rapid amplification，MIRA）技术建立快速检测SS及其血清2、7、9型3种主要临床血清型的新方法，该方法可在20~41 ℃温度下5~30 min内完成扩增，具有良好的特异性和敏感性；应用MIRA方法能够快速检测临床分离菌株中相应血清型，且与PCR方法的符合率为100%。上述结果表明，本研究所建立的MIRA方法可为临床诊断猪链球菌感染提供简便快捷的方法。

猪链球菌  /  血清型  /  多酶恒温快速扩增技术  /  快速检测

Streptococcus suis （SS） is one of the important pathogens causing serious economic losses in the pig farming industry. Here， we developed a novel rapid detection method for detecting SS and serovars 2， 7 and 9 based on multienzyme isothermal rapid amplification （MIRA） technology. The MIRA assays were validated for detection of SS and it serovars within 5 to 30 min at 20-41 ℃， with relative higher specificity and sensitivity. There are 100% agreement between our MIRA and PCR methods in the detection of clinical samples. Our results showed that the MIRA was a useful diagnostic method for rapid diagnosis of SS infection.

Streptococcus suis  /  serovars  /  MIRA  /  rapid detection
胡章玉, 张永青, 陈忆廉, 许发芝, 常馨月, 方富贵, 李亮, 孙裴, 刘雪兰. 基于多酶恒温快速扩增检测猪链球菌及其3种主要血清型方法的建立及应用. 畜牧兽医学报, 2026 , 57 (1) : 564 -568 . DOI: 10.11843/j.issn.0366-6964.2026.01.049
Zhangyu HU, Yongqing ZHANG, Yilian CHEN, Fazhi XU, Xinyue CHANG, Fugui FANG, Liang LI, Pei SUN, Xuelan LIU. Development and Evaluation of the Multienzyme Isothermal Rapid Amplification Assays for Detection of Streptococcus suis and Its Three Serovars[J]. Acta Veterinaria et Zootechnica Sinica, 2026 , 57 (1) : 564 -568 . DOI: 10.11843/j.issn.0366-6964.2026.01.049
正文
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猪链球菌（Streptococcus suis，SS）是影响全球养猪业的重要病原体之一，同时也是一种影响公共卫生的病原体。SS主要定植于猪的上呼吸道，尤其是扁桃体和鼻腔，或通过伤口感染，引起猪的急性败血症、脑膜炎、关节炎、心内膜炎、肺炎等疾病［1］，对养猪业造成严重影响。SS结构较复杂，血清型众多，临床上根据其荚膜多糖（capsular polysaccharide，CPS）基因的差异将SS分为29个血清型，因各个血清型之间存在的毒力因子不尽相同，从而导致临床上的发病症状也有所不同，其中SS2型对猪和人的致病性较强，影响公共卫生安全，此外，SS7和SS9型的感染在国内区域流行呈上升趋势［2⁃4］，这对控制猪链球菌病的发生带来了新的挑战，因此对猪链球菌感染的快速检测十分重要。
由于SS的生化反应差异较大，传统分离培养及生化鉴定技术难以将其准确分类，免疫学检测方法因相当比例的菌株不能定型，具有很大的不稳定性，容易出现交叉反应。目前，以聚合酶链式反应（polymerase chain reaction，PCR）技术为代表的分子生物学技术在临床应用最广泛［5］，多重PCR可以同时检测不同病原［6］，纳米PCR特异性强，操作方便，具有广阔的应用前景［7］，但上述方法均耗时长且操作复杂，依赖较为昂贵的仪器设备，因此，建立一种快速简便且对仪器设备要求不高的检测方法，对猪链球菌病防控具有重要意义。
多酶恒温快速扩增（multienzyme isothermal rapid amplification，MIRA）技术是一种新的等温扩增技术，在多种功能蛋白（解旋酶、单链结合蛋白、重组酶、DNA聚合酶）的协同作用下，于室温即可快速实现核酸扩增，对DNA模板不需要高温循环变性扩增，其反应过程始终在恒温下进行，对仪器的要求大大降低［8］。李盛杰和江海涛［9］建立了MIRA快速检测霍乱弧菌。本研究基于MIRA技术建立快速检测SS及其临床主要血清型2型（SS2）、7型（SS7）和9型（SS9）的方法，并将该方法应用于36份临床分离菌株的检测，为快速检测SS感染提供参考方法。
1　材料与方法
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1.1　菌株
猪链球菌、胸膜肺炎放线杆菌（Actinobacillus pleuropneumoniae，APP）、副猪嗜血杆菌（Haemophilus parasuis，HPS）、多杀性巴氏杆菌（Pasteurella multocida，Pm）、肠外致病性大肠杆菌（extraintestinal pathogenic E.coli，ExPEC）、肠炎沙门菌（Salmonella enteritidis，SE）临床分离菌株均为安徽农业大学病原微生物实验室保存。36株疑似SS菌株分离自病死猪的脾脏、肺脏、脑、关节液等，大部分菌株分离自安徽合肥、宿州和六安等，江苏盱眙、来龙和草堰等，少数来自广西贵港、山东泰安以及黑龙江绥化。
1.2　引物的合成与筛选
参考SS编码谷氨酸脱氢酶（glutamate dehydrogenase，gdh）基因以及NCBI上GenBank登录的SS2（BR001000.1）、SS7（BR001004.1）、SS9（BR001006.1）血清型的Cps基因的保守序列，通过Primer Premier 5.0软件设计MIRA扩增引物，序列均由安徽通用生物股份有限公司合成，引物信息见表1。
1.3　细菌基因组DNA的提取及MIRA反应体系的建立
按照细菌基因组DNA提取试剂盒（天根生化科技北京有限公司）操作步骤对细菌培养物进行基因组DNA的提取并通过Nanodrop进行核酸浓度的测定。参考DNA恒温快速扩增试剂盒（潍坊安普未来生物科技有限公司）步骤并稍作修改进行MIRA体系配制和扩增反应。在每个反应管中加入Buffer A（5.9 μL）、10 μmol上游引物（0.4 μL）、10 μmol下游引物（0.4 μL）、ddH2O和DNA模板（2.8 μL）、Buffer B（0.5 μL），充分混合并短暂离心后于37 ℃金属浴进行扩增反应。反应结束后，向每个反应管加入2.0 μL的6×Loading buffer，置于56 ℃孵育5 min，以2.0%琼脂糖凝胶电泳对扩增产物进行检测。
1.4　MIRA检测温度和时间的优化
设置反应温度分别为20、25、30、35、39、41 ℃，反应时间分别为5~30 min进行MIRA反应，分析扩增结果确定最佳反应温度和时间。
1.5　MIRA检测特异性和灵敏性
选用SS及各种血清型的最佳引物对，以SS种属和SS2、SS7、SS9血清型的基因组DNA为模板进行MIRA扩增反应，以APP、HPS、Pm、ExPEC、SE等临床常见猪细菌性病原DNA为对照，分析其特异性。再以10n梯度稀释的基因组DNA为模板进行MIRA扩增，分析其灵敏度。
1.6　临床分离菌株的MIRA检测
对36份SS临床分离菌株进行MIRA检测，并以PCR检测作为对照，比较两种方法的符合率。
2　结果
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2.1　MIRA引物的筛选
针对SS以及SS2、SS7与SS9血清型的MIRA引物筛选结果如图1所示，MIRA引物中SS-F2/R2、SS2-F2/R2、SS7-F3/R3和SS9-F3/R3具有相对较高的扩增效率和良好的特异性，可用于后续试验。
2.2　MIRA检测温度和时间的优化
优化MIRA检测反应条件的结果显示，在温度为20~41 ℃时，应用MIRA均可检测出SS及其SS2、SS7和SS9型，扩增速度快且效率较高的反应温度分别为30、35、30和35 ℃。应用MIRA扩增5 min即可检测出SS及SS2、SS7和SS9，但是目的条带相对较弱，延长反应时间至10 min以上可获得较为清晰的目的条带。由此可见，应用MIRA方法可快速检测SS及SS2、SS7和SS9。
2.3　MIRA检测的特异性分析
以猪源主要病原菌APP、HPS、Pm、ExPEC和SE等临床分离菌株为参考，用筛选出的MIRA引物对SS种属、SS2、SS7和SS9进行扩增。结果显示MIRA-SS、MIRA-SS2、MIRA-SS7和MIRA-SS9方法能够进行特异性检测，而与APP、HPS、Pm、ExPEC和SE等无明显交叉反应。显示了MIRA反应良好的特异性。
2.4　MIRA检测灵敏性分析
将SS（718 ng·μL-1）及SS2（718 ng·μL-1）、SS7（96 ng·μL-1）、SS9（191 ng·μL-1）基因组DNA进行100~10-7梯度稀释后进行MIRA扩增。结果显示，MIRA-SS和MIRA-SS2最低可检测7.18 pg·μL-1 DNA，而MIRA-SS7和MIRA-SS9最低检测限为0.96 pg·μL-1和1.91 pg·μL-1。
2.5　MIRA的临床分离株检测结果分析
为了验证MIRA在临床检测的应用，应用所建立的MIRA方法对36份临床分离的疑似SS菌株进行检测。结果显示，有30份呈SS阳性，其中2株为SS2型；1株为SS7型阳性：10株为SS9型，与PCR方法结果一致，两种方法的符合率均为100%（表2）。这表明所建立的MIRA检测SS以及SS2、SS7、SS9等方法可应用于SS临床分离菌株的检测。
3　讨论
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猪链球菌病是影响养猪业的一种重要的细菌病，死亡率可高达90%［10］，给养猪业造成了较大的经济损失。SS不同的血清型菌株中存在的主要毒力因子不同，导致患病动物发病的临床症状也不同，对能否及时采取应对措施带来了挑战。因此，迫切需要建立一种快速、有效和简便的检测方法。目前，临床上多采用PCR、环介导等温扩增（loop-mediated isothermal amplification，LAMP）等方法检测SS，如裴嘉琪［11］建立了多重PCR检测SS，最低检测限为100 pg·μL-1；刘博婷等［12］建立的LAMP方法检测SS2型时，相较于普通PCR检测方法，具有更高的准确性，但该方法也容易出现假阳性的情况。
MIRA方法是一种新兴的等温扩增技术，具有反应时间短、反应温度容易实现、操作简便等优点，由于不需要使用复杂的反应仪器，反应在常温下或水浴锅就可以进行，具有良好的应用前景。在近些年的研究中，MIRA广泛应用于检测各种物质，如Wang等［13］建立了MIRA快速检测海豚链球菌的方法并将MIRA方法与侧向流动试纸条相结合；Sun等［14］在将MIRA-LFD方法应用于检测乙型肝炎的临床样本时，在37 °C仅需10 min即可扩增出靶基因，反应灵敏度为10 pg，准确度为100%。Chen等［15］采用RT-MIRA检测将处理时间显著缩短至20 min；Hu等［16］在使用MIRA-LFD方法检测鲍曼不动杆菌时，将检测限降低至6 CFU·mL-1，显著优于RT-PCR，并且与其他弧菌无交叉反应，准确度为100%。
在本研究中建立的MIRA用于SS及其主要血清型的检测技术，可在反应温度为室温20 ℃即可检测到目的条带，表明本研究建立的MIRA方法在较低温度也可实现对SS的检测。在反应时间方面，MIRA仅反应5 min即可检测出SS，表明MIRA检测具有明显的即时性，能够节约反应时间。在特异性方面，本研究所建立的MIRA方法均能特异性检测出SS及相应血清型，与其他病原无交叉反应，且灵敏性分析结果显示，检测限低至7.18 pg·μL-1（SS2）、0.96 pg·μL-1（SS7）和1.91 pg·μL-1（SS9），具有良好的灵敏性。综上所述，本研究建立的MIRA方法可应用于临床快速检测SS及其临床主要血清型SS2、SS7、SS9，为防控猪链球菌病提供新的快速扩增技术。
4　结论
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本研究基于MIRA技术建立了可快速检测SS及SS2、SS7、SS9等临床血清型的新方法，该方法可在20~41 ℃温度下5~30 min内完成扩增，具有良好的特异性和敏感性，为临床防控SS提供快速诊断方法。
基金
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  • 安徽省高等学校科学研究重点项目(2022AH050898)
文献
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2026年第57卷第1期
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doi: 10.11843/j.issn.0366-6964.2026.01.049
  • 接收时间：2024-12-12
  • 首发时间：2026-09-16
  • 出版时间：2026-01-23
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  • 收稿日期：2024-12-12
基金
安徽省高等学校科学研究重点项目(2022AH050898)
作者信息
    安徽农业大学动物医学院，合肥 230036

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刘雪兰，主要从事病原微生物与免疫学研究，E-mail：
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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