Utility of automated real-time RT-PCR for the detection of foot-and-mouth disease virus excreted in milk
Utility of automated real-time RT-PCR for the detection of foot-and-mouth disease virus excreted in milk
复制标题
DOI：
10.1051/vetres:2005040
复制
发表时间：
2006-01-01
影响因子：
4.4
通讯作者：
Paton, DJ
中科院分区：
文献类型：
--
作者：
Reid, SM;Parida, S;Paton, DJ
口蹄疫病毒(FMDV)可在牛奶中排泄，从而将感染传播给其他持有者中的易感动物。通过研究实验感染牛的病毒排泄情况，评价了实时逆转录聚合酶链式反应(RRT-PCR)作为检测牛奶中口蹄疫病毒的诊断工具的可行性。从感染口蹄疫病毒的两头奶牛和同一围栏中的两头有接触的奶牛身上采集了4周的前奶和机奶样本。对牛奶的全脂、脱脂、奶油和细胞碎片成分进行了自动RRT-PCR检测，并与细胞培养中的病毒分离(VI)结果进行了比较。所有四头奶牛出现口蹄疫的临床症状都与病毒血症有关，在其他临床样本中也存在口蹄疫病毒。在检测所有牛奶成分中的口蹄疫病毒方面，RRT-PCR结果与VI结果非常一致，通常与临床症状的出现一致，但并不总是在临床症状出现之前。RRT-PCR在接种后23d的牛奶中检测到口蹄疫病毒，检出时间长于VI。此外，RRT-PCR检测牛奶中口蹄疫病毒的下限高于VI，与VI相反，RRT-PCR在模拟巴氏杀菌的热处理后检测到病毒基因组。RRT-PCR也能检测到防腐剂处理过的牛奶中的口蹄疫病毒。综上所述，本研究表明，自动化RRT-PCR比VI更快速、更敏感，可用于检测全脂牛奶和受感染动物的牛奶片段中的口蹄疫病毒。
Foot-and-mouth disease virus ( FMDV) can be excreted in milk and thereby spread infection to susceptible animals in other holdings. The feasibility of using real-time reverse transcription polymerase chain reaction (rRT-PCR) as a diagnostic tool for detection of FMDV in milk was assessed by studying the excretion of virus from experimentally-infected cattle. Fore- and machine milk samples were collected over a 4-week period from two dairy cows infected with FMDV and from two in-contact cows held in the same pen. The whole, skim, cream and cellular debris components of the milks were tested by automated rRT-PCR and results compared to virus isolation ( VI) in cell culture. The onset of clinical signs of FMD in all four cows correlated with viraemia, and the presence of FMDV in other clinical samples. rRT-PCR results matched closely with VI in detecting FMDV in all milk components and generally coincided with, but did not consistently precede, the onset of clinical signs. rRT-PCR detected FMDV in milk up to 23 days post inoculation which was longer than VI. Furthermore, the detection limit of FMDV in milk was greater by rRT-PCR than VI and, in contrast to VI, rRT-PCR detected virus genome following heat treatment that simulated pasteurisation. rRT-PCR was also able to detect FMDV in preservative-treated milk. In conclusion, this study showed that automated rRT-PCR is quicker and more sensitive than VI and can be used to detect FMDV in whole milk as well as milk fractions from infected animals.