Gonzague M, Plin C, Bakkali-Kassimi L, Boutrouille A, Cruciere C
AFSSA-Lerpaz, 22 rue Pierre Curie, BP67, 94703 Maisons-Alfort Cedex, France.
Mol Cell Probes. 2002 Jun;16(3):237-42. doi: 10.1006/mcpr.2002.0416.
For the detection of African swine fever virus (ASFV) by polymerase chain reaction (PCR) in clinical samples, an internal control was constructed to identify false negative results in each reaction. The internal control was designed in such a way that the same primer pair was used to amplify the internal control and the target DNA which were differentiated by size. The lower detection limit was reached at about 30 internal control DNA copies and about 50 genomic ASFV DNA copies. The use of the internal control revealed that from a total of 12 uninfected samples, 10 samples contained inhibitors. After a one in two dilution, all ten of these samples amplified the internal control satisfactorily. From a total of 16 samples from pigs suspected of having ASF infection, nine contained inhibitors. After a one in two dilution, we observed internal control amplification and target DNA amplification for four samples.
为了通过聚合酶链反应(PCR)在临床样本中检测非洲猪瘟病毒(ASFV),构建了一个内部对照以识别每个反应中的假阴性结果。内部对照的设计方式是使用相同的引物对扩增内部对照和目标DNA,二者通过大小进行区分。约30个内部对照DNA拷贝和约50个基因组ASFV DNA拷贝时达到较低检测限。使用内部对照发现,在总共12个未感染的样本中,有10个样本含有抑制剂。经过二分之一稀释后,所有这10个样本均能令人满意地扩增内部对照。在总共16个疑似感染ASF的猪的样本中,有9个含有抑制剂。经过二分之一稀释后,我们观察到4个样本的内部对照扩增和目标DNA扩增。