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

doi:10.11843/j.issn.0366-6964.2026.01.049

Abstract

Abstract:

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.

Key words: Streptococcus suis, serovars, MIRA, rapid detection

CLC Number:

S852.611

HU Zhangyu, ZHANG Yongqing, CHEN Yilian, XU Fazhi, CHANG Xinyue, FANG Fugui, LI Liang, SUN Pei, LIU Xuelan. 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.

Figures/Tables 3

Table 1

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Table 2

References 16

[1]	XU L，WANG X Y，LEI Z Y，et al.Spinal canal infection caused by Streptococcus suis in human：a case report［J］.BMC Infect Dis，2022，22（1）：369.
[2]	TAN M F，TAN J，ZHANG F F，et al.Exogenous glycogen utilization effects the transcriptome and pathogenicity of Streptococcus suis serotype 2［J］.Front Cell Infect Microbiol，2022，12：938286.
[3]	姜志康，韩先杰，郭皖露，等.猪链球菌7型的分离鉴定及致病性研究［J］.中国兽医科学，2025，55（2）：182-187.
	JIANG Z K，HAN X J，GUO W L，et al.Isolation，identification and pathogenicity of Streptococcus suis type 7［J］.Chinese Veterinary Science，2025，55（2）：182-187.（in Chinese）
[4]	陶攀，贾爱卿，张桂红，等.猪链球菌9型菌株的分离鉴定［J］.中国兽医杂志，2023，59（6）：93-96.
	TAO P，JIA A Q，ZHANG G H，et al.Isolation and identification of Streptococcus suis serotype 9 strain in swine［J］.Chinese Journal of Veterinary Medicine，2023，59（6）：93-96.（in Chinese）
[5]	SHUAI W，PARK B J，KIM S H，et al.Detection of Listeria monocytogenes using Dynabeads® anti-Listeria combined with real-time PCR in soybean sprouts［J］.LWT-Food Sci Technol，2019，99：533-539.
[6]	詹存林，王庚，孙秀秀，等.PRRSV、SS和Pm多重TaqMan荧光定量PCR检测方法的建立与初步应用［J］.中国兽医学报，2024，44（2）：231-236.
	ZHAN C L，WANG G，SUN X X，et al.Establishment and preliminary application of PRRSV，SS and Pm multiplex TaqMan real-time PCR detection methods［J］.Chinese Journal of Veterinary Science，2024，44（2）：231-236.（in Chinese）
[7]	张昆丽，林本夫，席振军，等.猪链球菌纳米PCR检测方法的建立和应用［J］.华北农学报，2021，36（4）：212-217.
	ZHANG K L，LIN B F，XI Z J，et al.Establishment of a nano PCR assay for detection of the Streptococcus suis［J］.Acta Agriculturae Boreali-Sinica，2021，36（4）：212-217.（in Chinese）
[8]	HENG P F，LIU J K，SONG Z，et al.Rapid detection of Staphylococcus aureus using a novel multienzyme isothermal rapid amplification technique［J］.Front Microbiol，2022，13：11027785.
[9]	李盛杰，江海涛.多酶恒温核酸快速扩增技术在霍乱弧菌检测中的应用［J］.南京晓庄学院学报，2021，37（6）：49-52.
	LI S J，JIANG H T.Application of multi-enzyme isothermal rapid amplification in the detection of Vibrio Cholera［J］.Journal of Nanjing Xiaozhuang University，2021，37（6）：49-52.（in Chinese）
[10]	杨风萍，董贵，黄文峰.猪链球菌病的防控［J］.养猪，2023（4）：92-95.
	YANG F P，DONG G，HUANG W F.Prevention and control of porcine streptococcal disease［J］.Swine Production，2023（4）：92-95.（in Chinese）
[11]	裴嘉琪.一种同时检测猪链球菌6个耐药基因的多重PCR方法的建立及初步应用［D］.武汉：华中农业大学，2023.
	PEI J Q.Establishment and preliminary application of a multiple PCR simultaneously detecting six resistance genes of Streptococcus suis［D］.Wuhan：Huazhong Agricultural University，2023.（in Chinese）
[12]	刘博婷，李娜，彭凌，等.副猪嗜血杆菌与猪链球菌2型双重LAMP检测方法的建立［J］.中国兽医科学，2018，48（9）：1080-1085.
	LIU B T，LI N，PENG L，et al.Development of duplex LAMP assay for the simultaneously detection of Haemophilus parasuis and Streptococcus suis type 2［J］.Chinese Veterinary Science，2018，48（9）：1080-1085.（in Chinese）
[13]	WANG Y F，NIU J J，SUN M M，et al.Rapid and sensitive detection of Streptococcus iniae in Trachinotus ovatus based on multienzyme isothermal rapid amplification［J］.Int J Mol Sci，2023，24（9）：7733.
[14]	SUN M L，LAI H Y，CHONG N Y，et al.Simple and feasible detection of hepatitis B virus via combination of multienzyme isothermal rapid amplification and lateral flow dipstick strip［J］.Front Mol Biosci，2021，8：763079.
[15]	CHEN H，ZHAO Z G，LIANG L，et al.Development of two novel on-site detection visualization methods for murine hepatitis virus based on the multienzyme isothermal rapid amplification［J］.Microb Pathog，2024，193：106776.
[16]	HU W W，HE J W，GUO S L，et al.Development and evaluation of a rapid and sensitive multienzyme isothermal rapid amplification with a lateral flow dipstick assay for detection of Acinetobacter baumannii in spiked blood specimens［J］.Front Cell Infect Microbiol，2022，12：1010201.

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
SS	SS-F1/R1	CAGAAACTGTCTTTATGCGATTGATT AGTCATTAAATTGTGGACGAGAGAGCT	427
	SS-F2/R2	GTAGGCTAACTAAAGCGGGCGATTGG ATTACGGGGTCATTTTGCCGAGTTCC	262
	SS-F3/R3	CCCATCCACCGTAAGACTAAGGTTTC CTTAGTTAGCCTACTGCGTCCCTCCA	156
SS2	SS2-F1/R1	GGGGTTCGTATTAACTTACTTGGCGTA TTAAGGGCATTTCAAGCGTTCCATTC	433
	SS2-F2/R2	ACTTTAGACTATGCTATAAGGGTTAGACGC TTCTACTGTTGCAGATAGGACTACAATGAT	222
	SS2-F3/R3	TTTAGGTTGGAATGGAACGCTTGAAATG CTCTACTGTTGCAGATAGGACTACAATG	188
SS7	SS7-F1/R1	TAGAGGTAGGTGGATTGCGTTCTTG AACACCCTGGATAGCCGTAATTGTA	215
	SS7-F2/R2	TAATAATGCCCTCGTGGAATACAGCTAAGT GGTGCCATAAATCATCTGAATCCAAGAACG	267
	SS7-F3/R3	TAGAGGTAGGTGGATTGCGTTCTTGG ATTCAAACACCCTGGATAGCCGTAATT	218
	SS7-F4/R4	TTAGTAGTAGGTGCTGGAGATGGTGGG ATGTTACCCGCCATAATGTCTTCAATGC	310
SS9	SS9-F1/R1	TATCAGACTGCATACAGGGCAGAAG ATCACAAGCAACCTGGGCAACAAAA	282
	SS9-F2/R2	TTGTCTCTCAAAATAAATCTAATCAGGGGC GAATTGTGAAACGACTTGCTACTAGAGGAT	490
	SS9-F3/R3	GGAAGTATTGGCAACGAAGAAGGGA CACATCATCAACAGCCTCTATATCCGC	177

Development and Evaluation of the Multienzyme Isothermal Rapid Amplification Assays for Detection of Streptococcus suis and Its Three Serovars

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