Detection of foot-and-mouth disease virus in milk samples by real-time reverse transcription polymerase chain reaction: Optimisation and evaluation of a high-throughput screening method with potential for disease surveillance.
Detection of foot-and-mouth disease virus in milk samples by real-time reverse transcription polymerase chain reaction: Optimisation and evaluation of a high-throughput screening method with potential for disease surveillance.
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通过实时逆转录聚合酶链反应来检测牛奶样品中的口和疾病病毒：优化和评估具有疾病监测潜力的高通量筛查方法。
DOI：
10.1016/j.vetmic.2018.07.024
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发表时间：
2018-09
影响因子：
3.3
通讯作者：
King DP
中科院分区：
文献类型：
--
作者：
Armson B;Mioulet V;Doel C;Madi M;Parida S;Lemire KA;Holder DJ;Das A;McIntosh MT;King DP
通过rRT-PCR在接触攻击后长达28天的牛奶中检测到FMDV。在从受感染农场采集的牛奶中检测到口蹄疫病毒（英国，2007年）。当牛奶样品在阴性牛奶中稀释至10−7时，FMDV检测是可能的。混合乳有可能成为FMDV监测的有价值的样品类型。本研究的目的是评估牛奶作为一种非侵入性样本类型的口蹄疫（FMD），一种高度传染性的偶蹄动物病毒性疾病的监测效用。4头泽西产奶奶牛通过与2头先前接种FMD病毒（FMDV：分离株O/UKG/34/2001）的非泽西产奶奶牛直接接触而感染。在整个感染过程中收集牛奶和血液，以比较两种高通量实时逆转录聚合酶链反应（rRT-PCR）方案与不同的RT-PCR化学。使用这两种方法，通过rRT-PCR在出现特征性足部病变前一到两天检测牛奶中的FMDV，与通过病毒分离检测相似。此外，与病毒分离检测（最多14 dpc）相比，从乳汁中进行的rRT-PCR检测延长至接触后28天（dpc）。此外，通过rRT-PCR检测牛奶中的FMDV可能比通过相同方法检测血清样品长18天。在2007年英国爆发期间收集的牛奶样本中也用两种rRT-PCR方法检测到FMDV。稀释研究使用来自田间和实验感染动物的牛奶进行，其中对于一个样品，可以在10−7检测到FMDV。根据本研究中检测到的CT峰值，这些结果表明，使用散装罐或奶罐车中的牛奶，有可能在典型规模的奶牛群（100-1000头）中识别出一头急性感染的奶牛。这些结果促使进一步研究在口蹄疫流行国家使用牛奶进行口蹄疫监测。
FMDV was detected by rRT-PCR in milk up to 28 days post contact challenge. FMDV was detected in milk collected from infected farms in the field (UK 2007). FMDV detection was possible when a milk sample was diluted up to 10−7 in negative milk. Pooled milk has the potential to be a valuable sample type for FMDV surveillance. This study aimed to evaluate the utility of milk as a non-invasive sample type for the surveillance of foot-and-mouth disease (FMD), a highly contagious viral disease of cloven-hooved animals. Four milking Jersey cows were infected via direct-contact with two non-milking Jersey cows that had been previously inoculated with FMD virus (FMDV: isolate O/UKG/34/2001). Milk and blood were collected throughout the course of infection to compare two high-throughput real-time reverse transcription polymerase chain reaction (rRT-PCR) protocols with different RT-PCR chemistries. Using both methods, FMDV was detected in milk by rRT-PCR one to two days before the presentation of characteristic foot lesions, similar to detection by virus isolation. Furthermore, rRT-PCR detection from milk was extended, up to 28 days post contact (dpc), compared to detection by virus isolation (up to 14 dpc). Additionally, the detection of FMDV in milk by rRT-PCR was possible for 18 days longer than detection by the same method in serum samples. FMDV was also detected with both rRT-PCR methods in milk samples collected during the UK 2007 outbreak. Dilution studies were undertaken using milk from the field and experimentally-infected animals, where for one sample it was possible to detect FMDV at 10−7. Based on the peak CT values detected in this study, these findings indicate that it could be possible to identify one acutely-infected milking cow in a typical-sized dairy herd (100–1000 individuals) using milk from bulk tanks or milk tankers. These results motivate further studies using milk in FMD-endemic countries for FMD surveillance.
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DOI：
10.2460/javma.2002.220.1636
发表时间：
2002-06-01
期刊：
JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION
影响因子：
--
作者：
Callahan, JD;Brown, F;Nelson, WM
通讯作者：
Nelson, WM
影响因子：
3.1
作者：
Reid, SM;Forsyth, MA;Ferris, NP
通讯作者：
Ferris, NP
影响因子：
1.5
作者：
King, DP;Ferris, NP;Beckham, TR
通讯作者：
Beckham, TR
影响因子：
1.5
作者：
Renshaw, RW;Ray, R;Dubovi, EJ
通讯作者：
Dubovi, EJ
DOI：
10.20506/rst.21.3.1343
发表时间：
2002-12-01
影响因子：
0.6
作者：
Kitching, RP
通讯作者：
Kitching, RP