Establishment and Application of a Multiplex RT-PCR Assay for Detection of PEDV,TGEV and PRoV     

LONG Fei-xiang  SHI Kai-chuang  ZHANG Zhen  YIN Yan-wen  CHEN Han-zhong  MO Sheng-lan 《中国畜牧兽医》2016,43(10):2518-2526

In this study,a multiplex RT-PCR assay was established to differentially detect porcine epidemic diarrhea virus (PEDV),porcine transmissible gastroenteritis virus (TGEV) and porcine rotavirus (PRoV) after optimization of the reaction conditions.Three pairs of primers PEDV-N,TGEV-M and PRoV-VP6 were designed for specifically amplifying PEDV N gene,TGEV M gene and PRoV VP6 gene,respectively.The assay could specifically amplify PEDV,TGEV and PRoV,but not classical swine fever virus (CSFV),porcine foot and mouth disease virus (FMDV),pseudorabies virus (PRV),porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2).The detection limits of PEDV,TGEV and PRoV standard recombinant plasmids were 1.41×10³,1.41×10² and 1.41×10³ copies/μL,respectively.The repeated reaction under the same conditions obtained uniform results.The assay was used to detect a total number of 190 clinical samples,of which 42 (22.11%) samples were positive for PEDV,58 (30.53%) samples for TGEV and 34 (17.89%) samples for PRoV,and there were mixed infection among these viruses.The results indicated that this multiplex RT-PCR assay had the advantages of sensitivity,specificity and repeatability and provided a useful tool for differential detection and epidemiological investigation of PEDV,TGEV and PRoV.  相似文献   

Study on Expression Difference of IGFBP-5 Gene in Different Tissues of Kazakh and Yanqi Horses     

TUERXUNJIANG&#  Wumueraili  MENG Jun  WANG Jian-wen  ZENG Ya-qi  YAO Xin-kui  LI Lin-ling  YAO Fang-fang  WANG Huan  REN Hai-fan  HUANG Jing-jing  A Mi-na 《中国畜牧兽医》2016,43(9):2257-2264

The study aimed to explore the mRNA expression pattern of insulin-like growth factor binding protein-5 (IGFBP-5) gene in different tissues of Kazakh and Yanqi horses.The expression of IGFBP-5 gene in different tissues of heart,liver,spleen,lung,kidney,small intestine,large intestine,cecum,intercostal muscles,longissimus dorsi muscles,brachialis muscle and gluteus in two horses were detected by Real-time quantitative PCR and compared the mRNA expression in the same tissues of two breeds.The results showed that the expression of IGFBP-5 in longissimus dorsi muscle and brachialis muscle of two breeds were significantly higher than other tissues including heart,liver,spleen,lung,kidney,small intestine,large intestine and cecum (P<0.05),and was the lowest in large intestine.The expression of IGFBP-5 in kidney,small intestine,large intestine,cecum,longissimus dorsi muscle and brachialis muscles of Yanqi horse were higher than in the same part of Kazakh horse,and among those in longissimus dorsi muscle and large intestine of Yanqi horse were extremely significantly higher than in the same part of Kazakh horse (P<0.01),and in small intestine and cecum of Yanqi horse were significantly higher than in the same part of Kazakh horse (P<0.05).The test was for further researching the biological function of IGFBP-5 gene,and it could provide a theoretical basis for genetic improvement of production performance of horse in our country.  相似文献   

Establishment and Application of Nested RT-PCR Assay for Detection of Classical Swine Fever Virus     

LIU Mei-fen  WANG Shu-juan  YAN Ruo-qian  WANG Dong-fang  CAO Wei-wei  ZHAO Xue-li  LI Qin-nan  LI Ning 《中国畜牧兽医》2016,43(9):2285-2290

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Development of A Dual Real-time PCR for the Rapid Detection of Listeria monocytogenes     

LI Dan-dan  XU Yi-gang  LI Meng-yuan  WANG Yu  QIU Suo-ping  GAO Hui-jiang  GAO Shen-yang 《中国畜牧兽医》2016,43(6):1453-1457

To establish a rapid assay for Listeria monocytogenes(LM) detection,a Real-time PCR method was developed targeting iap gene of LM.The results showed that the test for 15 bacteria strains,only LM was positive,indicated that the method had high specificity.In addition,the sensitivity of Real-time PCR was 6.5 CFU/mL.Stability and reproducibility of the test showed that the coefficient of variation for the same sample repeat the Ct values were less than 2%.Furthermore,a total of 3 positive samples for LM were detected from 139 clinical samples by the method,which was in accordance with the testing result by GB 478930-2010 standard detection protocol.Therefore,the Real-time PCR method provides a novel rapid,sensitive and good repeatability detection method for LM infection.  相似文献   

Expression Analysis of THBS3 Gene in Different Tissues and Skeletal Muscles at Different Periods from Landrace and Tongcheng Pigs     

MA Xi-shan  TANG Zhong-lin  XIAO Chong  YANG Bo-chao  QIN Chuan 《中国畜牧兽医》2016,43(4):1032-1038

To study the expression pattern of THBS3 gene in different tissues and during skeletal muscle development, the THBS3 gene expression in different tissues and skeletal muscles during prenatal periods (33, 45, 65, 70 and 90 d) and postnatal periods (0, 9, 30, 60, 120 and 160 d) from Landrace and Tongcheng pigs were detected by Real-time quantification PCR.The results showed that THBS3 gene widely expressed in all tissues examined, exhibiting similar spatial expression patterns with expression peaks in lung in both pig breeds except in stomach and intestine.Moreover, although THBS3 gene showed a significant higher expression level in gestation than after birth in Landrace and Tongcheng pigs (P<0.05), it exhibited different expression patterns between Landrace and Tongcheng pigs, the expression peak was detected at gestation day 45 in Landrace pig, while was detected at gestation day 65 in Tongcheng pig.The results suggested that THBS3 gene involved in skeletal muscle growth and development in pigs, as well as the regulation of asynchronization of skeletal muscle development in different pig breeds.  相似文献   

Establishment of a Duplex Real-time RT-PCR Assay for the Differential Detection of Nipha Virus and Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus     

WANG Jian-hua  DONG Zhi-zhen  WANG Yu-ling  XIAO Yan  ZHAO Xiang-ping 《中国畜牧兽医》2016,43(2):348-355

To establish a rapid,sensitive and specific assay for the differential detection of Nipah virus (NiV) and highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV),a duplex Real-time RT-PCR was developed with specific primers and probes targeting to the special sequences of NiV M gene and HP-PRRSV nsp2 gene by optimization of reaction conditions.The performance of the assay was linear ranging from 4.6×10¹ to 4.6×10⁷ copies/μL for RNA standard control of NiV M (NiV-M-RNA) and from 4.1×10¹ to 4.1×10⁸ copies/μL for RNA standard control of HP-PRRSV nsp2 (HP-PRRSV-nsp2-RNA),and detection limits of the assay was 46 copies for the NiV-M-RNA and 4.1 copies for the HP-PRRSV-nsp2-RNA,respectively.The coefficients of variation (CVs) of both inter-assay and intra-assay repeatability were less than 2.0%,showing good repeatability.The assay was able to specifically detect NiV and HP-PRRSV simultaneously without cross-reaction with classical swine fever virus (CSFV),porcine epidemic diarrhea virus (PEDV),swine influenza virus (SIV),porcine parvovirus (PPV),pseudorabies virus (PRV) and porcine circovirus type 2 (PCV2).Of the 236 samples from pigs for both NiV and HP-PRRSV detection by the established assay,all the samples were negative for NiV,8 samples were HP-PRRSV positive.In conclusion,this assay offers a useful approach for the differential detection of NiV and HP-PRRSV in clinical specimens from the pigs.  相似文献   

Analysis on Polymorphisms of Three CNV Regions in Five Pig Breeds     

HUANG Shi-hui  RAN Xue-qin  WANG Jia-fu  PAN Hua  LI Rong-rong 《中国畜牧兽医》2016,43(3):730-737

The paper was aimed to investigate the polymorphism of copy number variation (CNV) in different pig breeds.Three CNV regions of CNVR91,CNVR92 and CNVR143 were chosen from the porcine SNP60 chip genotyping results.The polymorphisms of three CNVs were determined by Real-time quantitative PCR method,taking five pig breeds as samples,including Yorkshire pig,Xiang pig,Kele pig,Nuogu pig and Rongchang pig breeds.The results showed that the dominant status of CNVR91 was loss in Xiang pig,while it was normal in other four pig breeds.The major type of CNVR92 was deletion in Xiang pig,Yorkshire pig,Kele pig and Rongchang pig breeds,with a high normal percent in Nuogu pig.For CNVR143,the dominant event was gain in Xiang pig and Nuogu pig breeds,but it was not diverse in other three pig breeds.These results indicated that three CNV regions emerged with polymorphism in five pig breeds,which might have effects on gene expression in CNV regions and physiological function by dosage effect especially in Xiang pig,Nuogu pig and Kele pig breeds.  相似文献   

Confirmation of sex-specific transcripts from Ancylostoma caninum adult worms by semi-quantitative RT-PCR     

Miranda RR  Clara e Silva L  Santos HA  Rabelo EM 《Research in veterinary science》2007,82(2):215-217

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基于冠层温度和土壤墒情的实时监测与灌溉决策系统   总被引：4，自引：0，他引：4  

蔡甲冰  许迪  司南  魏征 《农业机械学报》2015,46(12):133-139

设计了一个可以在线连续监测田间作物冠层温度、环境信息和土壤墒情的实时灌溉决策系统,并将其安装于农田进行了1 a实际运行和观测。系统采用太阳能供电和微处理器进行数据采集和管理,为野外的实际应用提供了保障。系统配置了红外温度、空气温/湿度、土壤水分/水势等传感器,能够及时采集田间全面的同步数据,排除了异地观测所形成的数据误差。采用悬臂式多点采集下垫面红外温度检测方法,可以快速采集更多和更高精度的数据,避免单点测量的人为误差。系统配备的快速锁紧装置,能够根据下垫面作物的生长情况进行传感器位置高度调节,使检测数据更符合田间实际情况。通过运行管理和监测数据分析可见,所监测数据能够很精细的刻画田间作物实际生长状况,可以用于灌区综合灌溉决策,实现田间精量灌溉管理和控制,为灌溉管理的精量化和智能化提供数据支持。  相似文献   

神经肽S及其受体在猪免疫器官中的表达及分布   总被引：1，自引：0，他引：1  

林小琴  姚远  周莹珊  闭璟珊  崔金鑫  周春燕  雷治海 《畜牧与兽医》2010,42(8)

本研究采用半定量RT-PCR方法,对神经肽S(NPS)及其受体(NPSR)mRNA在猪免疫器官中的表达水平进行检测。结果表明,NPSmRNA在胸腺、脾、空肠淋巴结和软腭扁桃体中均有表达,且在脾和胸腺中的表达水平较高;NPSR mRNA在脾脏的表达水平最高。进一步利用免疫组织化学法对NPS在猪免疫器官中的分布定位进行了研究,在猪脾脏和空肠淋巴结均有NPS免疫阳性细胞分布,但在软腭扁桃体和胸腺中未见NPS免疫阳性细胞。再进行免疫接种猪传染性胃肠炎疫苗,运用半定量RT-PCR技术检测免疫接种后不同时间(3、7、14 d)NPS及其受体mRNA在猪免疫器官中表达水平的变化规律。结果表明,NPS mRNA和NPSR mRNA在胸腺、脾、淋巴结和扁桃体中均有表达,且在一定时间范围内呈现先升高后下降的变化规律,但对照组无明显的升高和下降现象。上述结果提示,猪NPS可能参与免疫功能的调节。  相似文献