副溶血弧菌实时荧光定量PCR快速检测方法的建立

doi:10.16431/j.cnki.1671-7236.2016.01.009

Abstract

Abstract: To establish a rapid assay for detection of Vibrio parahaemolyticus(VP), Real-time PCR method was developed targeting to toxR gene of Vibrio parahaemolyticus.The results showed that the test for 15 bacteria strains using the Real-time PCR method, only Vibrio parahaemolyticus test was positive, indicating that the method had high specificity.In addition, the sensitivity of Real-time PCR was 4.9 CFU/mL.Furthermore, a total of 3 positive samples for Vibrio parahaemolyticus were detected from 150 clinical samples by the Real-time method, which was in accordance with the testing result by GB 4789.7-2013 standard detection protocol.Therefore, the Real-time method provided a novel rapid and sensitive detection method with good practicality for Vibrio parahaemolyticus infection.

Key words: Vibrio parahaemolyticus(VP); toxR gene; Real-time PCR

CLC Number:

LI Dan-dan, XU Yi-gang, WANG Yu, QIU Suo-ping, GAO Hui-jiang, GAO Shen-yang. Development of Real-time PCR Method for Rapid Detection of Vibrio parahaemolyticus[J]. , 2016, 43(1): 58-62.

References

[1] 林强,李宁求,付小哲,等.牡蛎中副溶血弧菌荧光定量PCR检测方法的建立及其应用[J].中国水产科学,2011,18(1):96-102.
[2] 纪懿芳,胡文忠,姜爱丽,等.海产品中副溶血弧菌检测方法研究进展[J].食品工业科技,2015,36(5):365-369.
[3] Altekruse S F,Bishop R D,Baldy L M,et al. Vibrio gastroenteritis in the US Gulf of Mexico region:The role of raw Ostrea[J]. Epidemiol Infect,2000,124(3):489-495.
[4] 徐义刚,李丹丹,刘新亮,等.基于环介导等温扩增技术水产品中副溶血弧菌快速检测方法的建立与应用[J].中国兽医杂志,2013,49(10):68-71.
[5] 宫春波,王朝霞,刘磊,等.烟台市生食水产品中副溶血弧菌污染调查及风险评价[J].食品安全质量检测学报,2014,5(11):3758-3763.
[6] 段永翔,鞠长燕,刘李,等.2006-2008年深圳市食源性病例和外环境的副溶血性弧菌分离株分型特征比较[J].微生物学报,2011,51(4):518-523.
[7] Gooch J A,DePaola A,Kaysner C A,et al.Evaluation of two direct plating methods using nonradioactive probes for enumeration of Vibrio parahaemolyticus in oysters[J]. Appl Environ Microbiol,2001,67(2):721-724.
[8] 张晓君,梁利国,阎斌伦,等.基于toxR基因的SYBR GreenⅠ实时定量PCR检测病原副溶血弧菌方法的建立[J].中国兽医学报,2012,32(2):543-547.
[9] 邵美丽,董鑫,赵燕丽,等.单增李斯特菌和金黄色葡萄球菌双重荧光定量PCR检测方法建立[J].食品科学,2013,34(16):169-172.
[10] 李丹丹,徐义刚,王绥家,等.肠出血性大肠埃希菌O157:H7实时荧光定量PCR快速检测方法的建立[J].中国畜牧兽医,2014,41(7):81-84.
[11] 刘二龙,卢丽,唐婕,等.化脓隐秘杆菌TaqMan实时荧光定量PCR检测方法的建立[J].中国畜牧兽医,2015,42(5):1093-1097.
[12] Cai T,Jiang L,Yang C,et al.Application of Real-time PCR for quantitative detection of Vibrio parahaemolyticus from seafood in eastern China[J]. FEMS Immunol Med Microbiol,2006,46(2):180.
[13] 冯可,胡文忠,姜爱丽,等.单核细胞增生性李斯特菌分子生物学检测技术的研究进展[J].2014,35(4):392-396.
[14] 范斌,许文花,韩建强,等.猪瘟兔化弱毒疫苗中病毒含量实时荧光定量RT-PCR检测方法的建立及初步应用[J].中国畜牧兽医,2014,41(12):56-61.
[15] 倪梦丽,杨冰,王春德,等.贝类中副溶血弧菌和沙门氏菌荧光定量PCR快速检测方法的建立[J].中国动物检疫,2013,30(8):78-82.
[16] 冼钰茵,易敏英,魏霜,等.基于弧菌毒力基因多重PCR法的建立及其在副溶血弧菌毒力基因检测中的应用[J].现代食品科技,2015,31(7):309-315.

Development of Real-time PCR Method for Rapid Detection of Vibrio parahaemolyticus

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	MA Baoyi, SHENG Chenyan, LIU Hongying, MA Qingxia, WANG Tingting, LI Jianming, SHI Kun, DU Rui, LENG Xue. Establishment of a Duplex TaqMan Real-time PCR Method for Detection of Bovine Viral Diarrhea Virus Types 1 and 2 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2326-2335.
[2]	LI Jianmei, JI Zhengjian, LIU Mei, SHEN Xinyue, CHENG Xu, YU Yan, DAI Yabin. Expression Changes of Immune-related Genes in Tibetan Chickens and Recessive White Chickens Following Eimeria tenella Infection [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1440-1448.
[3]	JIANG Hui, LIU Pan, LI Yuanyuan, LIU Kuiguo, HUANG Xixia, LIU Jiangwei, XU Lei, DU Jianwen, SANG Zhagen, YANG Xuemei, ZHAO Fanfan, WANG Dan. Expression and Bioinformatics Analysis of Mastitis Resistance Related Genes in Xinjiang Brown Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(1): 90-97.
[4]	XU Min, XU Houqiang, CHEN Wei, YANG Yang. Study on Expression of the Relate Genes Involved in Intramuscular Preadipocyte Differentiation in Congjiang Xiang Pig [J]. , 2018, 45(9): 2492-2499.
[5]	ZHAO Xueli, LIU Yi, BAN Fuguo, YAN Ruoqian, WANG Huajun, WANG Shujuan, MA Zhenyuan, WANG Dongfang. Establishment and Application of A Double TaqMan MGB Real-time PCR Assay for Detection of Canine Distemper Virus and Canine Parvovirus [J]. , 2018, 45(8): 2086-2094.
[6]	LAN Desong, GU Guibo, HOU Zhengzhong. Development of a TaqMan Real-time PCR for Differentiation of Wild-type Strain from gE-deleted Vaccine Strain of Pseudorabies Virus [J]. , 2018, 45(7): 1804-1812.
[7]	GAO Xintao, JIA Hong, HOU Shaohua, GUO Xiaoyu, YUAN Weifeng, JIANG Yitong, ZHU Hongfei, XIN Ting. Research on the Potential of IL-17 as a Biomarker of Bovine Tuberculosis and Establishment of Its Real-time PCR Assay [J]. , 2018, 45(6): 1447-1453.
[8]	MAO Ning, WANG Jiafu, ZHANG Fuping, TANG Liangting, RUAN Yiqi, YI Fanli, RAN Xueqin. Study on Differential Expression of StAR and CYP11A1 Genes in Xiang Pig Ovary [J]. , 2018, 45(5): 1137-1144.
[9]	WAN Chunhe, LIU Rongchang, CHENG Longfei, SHI Shaohua, FU Guanghua, CHEN Hongmei, FU Qiuling, HUANG Yu. Establishment of a TaqMan-based Real-time PCR Method of Duck Adenovirus A [J]. , 2018, 45(5): 1341-1348.
[10]	ZHANG Mengyao, YANG Feng, LIU Kaidong, CHENG Ming, WANG Guoyi, LIU Jifeng, LIU Nan, HE Jianning. Study on Construction of DKK1 Gene Recombinant Plasmid in Aohan Fine Wool Sheep and Its Expression in Fibroblasts [J]. , 2018, 45(1): 131-139.
[11]	BU Xiao-yan, LI Hui, SHI Xiao-li, SHI Xin-cai, ZHAO Zhong-hai, CHEN Lin. Study on Different Expression of HO-1 Gene of 70-weeks-old Changshun Blue-shelled Chicken in Different Tissues [J]. , 2017, 44(6): 1762-1768.
[12]	LI Yuan-ji, YANG Ming-hua, LIU Fang, ZHU Fang-fang, HE Shao-yu, KUANG Hai-ou, LI Ya-hui. cDNA Cloning, Tissue Expression Profiles and Bioinformatical Analysis of Tssk6 Gene in Tree Shrews [J]. , 2017, 44(3): 619-627.
[13]	CAI Wen-yi, CHEN Tong, SHI Xian, CHEN Wei-ming, HU Jia-jia, XIONG Xian-rong, LAN Dao-liang, LI Jian. Cloning of PLIN2 Gene of Yak and Its Expression in Ovary during Different Estrus Periods [J]. , 2017, 44(3): 782-789.
[14]	HU Feng-li, XU Li-yuan, ZHANG Zhi-fei, ZHOU Wei-guang. Screening of siRNA Targeted for TLR9 Gene and Its Effect on the Proliferation Profile of BHV-1 [J]. , 2017, 44(2): 303-310.
[15]	HE Shuai, ZHOU Chun-xue, ZHU Xing-quan, ZHOU Dong-hui, XU Qian-ming. Study on Expression of Toxoplasma gondii Rhoptry Neck Protein 5 Gene in Different Developmental Stages [J]. , 2017, 44(2): 350-356.