A quadruplex real-time PCR assay combined with a conventional PCR for the differential detection of Marek's disease virus vaccines and field strains.
A quadruplex real-time PCR assay combined with a conventional PCR for the differential detection of Marek's disease virus vaccines and field strains.
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DOI：
10.3389/fvets.2023.1161441
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发表时间：
2023
影响因子：
3.2
通讯作者：
中科院分区：
文献类型：
--
作者：
为了评价疫苗的免疫效果，并区分疫苗与MDV强毒，建立了一种新的基于TaqMan探针的四重实时荧光定量PCR方法，用于区分和准确定量HVT、CVI 988和MDV-1强毒。结果表明，该方法对CVI 988、HVT和MDV强毒DNA的检测限均为10拷贝，相关系数均大于0.994，与其它禽病病毒无交叉反应。新检测试剂的Ct值的批内和批间变异系数（CV）均小于3%。对CVI 988和MDV强毒在感染后7 ~ 60 d收集的羽毛中的复制动力学分析表明，MD5对CVI 988的基因组载量没有显著影响（p > 0.05），而接种CVI 988可显著降低MD5的病毒载量（p < 0.05）。与meq基因PCR技术相结合，可有效地鉴定MDV强毒感染。结果表明，该方法能有效区分疫苗和MDV强毒株，具有可靠、灵敏、特异的优点，可用于MDV强毒株免疫状态的确认和传播监测。
To evaluate the effect of the vaccine and differentiate vaccine from virulent MDV, a new quadruplex real-time PCR assay based on TaqMan probes was developed to differentiate and accurately quantify HVT, CVI988 and virulent MDV-1. The results showed that the limit of detection (LOD) of the new assay was 10 copies with correlation coefficients >0.994 of CVI988, HVT and virulent MDV DNA molecules without cross-reactivity with other avian disease viruses. The intra-assay and inter-assay coefficients of variation (CVs) of Ct values for the new assay were less than 3%. Analysis of replication kinetics of CVI988 and virulent MDV of collected feathers between 7 and 60 days post-infection (dpi) showed MD5 had no significant effect on the genomic load of CVI988 (p > 0.05), while vaccination with CVI988 could significantly reduce the viral load of MD5 (p < 0.05). Combined with meq gene PCR, this method can effectively identify virulent MDV infections in immunized chickens. These results demonstrated that this assay could distinguish between the vaccine and virulent MDV strains and had the advantages of being reliable, sensitive and specific to confirm the immunization status and monitor the circulation of virulent MDV strains.
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DOI：
10.1073/pnas.0404508101
发表时间：
2004-08-10
影响因子：
11.1
作者：
Lupiani, B;Lee, LF;Reddy, SM
通讯作者：
Reddy, SM
影响因子：
4.8
作者：
Conradie, Andele M.;Bertzbach, Luca D.;Kaufer, Benedikt B.
通讯作者：
Kaufer, Benedikt B.
影响因子：
2.8
作者：
Handberg, KJ;Nielsen, OL;Jorgensen, PH
通讯作者：
Jorgensen, PH
DOI：
10.1016/s1872-2075(08)60028-x
发表时间：
2008-04-01
期刊：
Sheng wu gong cheng xue bao = Chinese journal of biotechnology
影响因子：
--
作者：
Cui, Hongyu;Wang, Yunfeng;Wang, Mei
通讯作者：
Wang, Mei
影响因子：
1.6
作者：
Spatz, Stephen J.;Schat, Karel A.
通讯作者：
Schat, Karel A.