[ Objective] This study aimed to establish a rapid, sensitive and specific SYBR Green I real-time quantitative PCR assay for the detection of latent pseudorabies virus （PRV） infection. [ Method ] SYBR Green I real-t...[ Objective] This study aimed to establish a rapid, sensitive and specific SYBR Green I real-time quantitative PCR assay for the detection of latent pseudorabies virus （PRV） infection. [ Method ] SYBR Green I real-time quantitative PCR conditions and system for early detection of latent pseudorabies virus in- fection were optimized and compared with conventional PCR to investigate the sensitivity and specificity. Subsequently, the established assay was applied to detect different clinical samples. [ Result] The sensitivity of SYBR Green I real-time quantitative PCR assay （52 copies/μl） was 1 000 times higher than that of conven- tional PCR （5.2×1^04 copies/μl） and the detection time was shortened by 1/2. The established assay could be used to detect PRV but could not be used to detect PCV2, PPV, CSFV or PRRSV. Various tissues were collected from Bama miniature pigs with latent PRV infection under sterile conditions for real-time PCR detection. Results showed that viral copy number in the brain, nasal swab, inguinal lymph node, liver, lung and spleen was above 20, while PRV was not detected in the kidney and heart tissues. [ Conclusion] The established SYBR Green I real-time quantitative PCR assay for PRV/.AT detection was specific, sensitive and rapid, which could be used for pathogen monitoring, epidemiological investigation and quantitative study of PRV.展开更多
Recently, antibiotics have been widely used to resist various bacterial diseases in livestock breeding, which generated more superbacteria with resistance. Food safety problems, such as animal drug residues are urgent...Recently, antibiotics have been widely used to resist various bacterial diseases in livestock breeding, which generated more superbacteria with resistance. Food safety problems, such as animal drug residues are urgent to be solved. Traditional Chinese medicines are traditional treasures of China, which attract more and more attention in the prevention and treatment of bacterial diseases because of their abundant advantages including improving immunologic function of or-ganisms, antibacterial, antivirus, antitumor, anti-inflammatory and improving animal performance. The advantages of traditional Chinese medicines, such as low toxicity, low residual, green and safe, conform to food safety requirements. The using condition of traditional Chinese medicines in main bacterial diseases of animals was reviewed in the paper, thus to provide a reference for livestock production.展开更多
基金Supported by Shandong Provincial Natural Science Foundation(ZR2012CQ012)Shandong Provincial Technical Innovation Grant(201220916006)Special Fund for Applied Technology Research and Development of Binzhou City(200706)
文摘[ Objective] This study aimed to establish a rapid, sensitive and specific SYBR Green I real-time quantitative PCR assay for the detection of latent pseudorabies virus （PRV） infection. [ Method ] SYBR Green I real-time quantitative PCR conditions and system for early detection of latent pseudorabies virus in- fection were optimized and compared with conventional PCR to investigate the sensitivity and specificity. Subsequently, the established assay was applied to detect different clinical samples. [ Result] The sensitivity of SYBR Green I real-time quantitative PCR assay （52 copies/μl） was 1 000 times higher than that of conven- tional PCR （5.2×1^04 copies/μl） and the detection time was shortened by 1/2. The established assay could be used to detect PRV but could not be used to detect PCV2, PPV, CSFV or PRRSV. Various tissues were collected from Bama miniature pigs with latent PRV infection under sterile conditions for real-time PCR detection. Results showed that viral copy number in the brain, nasal swab, inguinal lymph node, liver, lung and spleen was above 20, while PRV was not detected in the kidney and heart tissues. [ Conclusion] The established SYBR Green I real-time quantitative PCR assay for PRV/.AT detection was specific, sensitive and rapid, which could be used for pathogen monitoring, epidemiological investigation and quantitative study of PRV.
基金Supported by Natural Science Foundation of Shandong Province(ZR2012CQ012)Shandong Province Modern Agricultural Technology System of Swine Innovation Team(SDAIT-08-17)
文摘Recently, antibiotics have been widely used to resist various bacterial diseases in livestock breeding, which generated more superbacteria with resistance. Food safety problems, such as animal drug residues are urgent to be solved. Traditional Chinese medicines are traditional treasures of China, which attract more and more attention in the prevention and treatment of bacterial diseases because of their abundant advantages including improving immunologic function of or-ganisms, antibacterial, antivirus, antitumor, anti-inflammatory and improving animal performance. The advantages of traditional Chinese medicines, such as low toxicity, low residual, green and safe, conform to food safety requirements. The using condition of traditional Chinese medicines in main bacterial diseases of animals was reviewed in the paper, thus to provide a reference for livestock production.
