Bogdanovic G, Brytting M, Cinque P, Grandien M, Fridell E, Ljungman P, Lönnqvist B, Hammarin A L
Swedish Institute for Infectious Disease Control, Stockholm, Sweden.
Clin Diagn Virol. 1994 Apr;2(2):127-36. doi: 10.1016/0928-0197(94)90045-0.
A nested polymerase chain reaction (PCR) was developed to detect BK virus (BKV) and JC virus (JCV) DNA sequences. The unique clevage site for BamHI restriction enzyme was located in teh JCV amplimer and cleavage was used to differentiate between BKV and JCV.
Twenty-three urine specimens from 17 bone marrow recipients with haemorrhagic cystitis and one liver transplant patient were tested for the presence of BKV and JCV DNA. Four brain tissue specimens (paraffin embedded brain tissues and a fresh frozen brain biopsy) and 5 cerebrospinal fluids from 3 AIDS patients and one liver transplant patient, all with progressive multifocal leukoencephalopathy (PML), were also examined by PCR.
The sensitivity of the PCR was 10 genomes for each virus. BKV DNA was detected in 15 urine specimens from 12 bone marrow transplant patients. JCV DNA was detected in 4 cerebrospinal fluids and 4 brain tissues from patients with PML.
Our results show that the nested PCR is a sensitive and rapid assay that can be used for diagnosis of BKV and JCV infections.
开发了一种巢式聚合酶链反应(PCR)来检测BK病毒(BKV)和JC病毒(JCV)的DNA序列。BamHI限制性内切酶的独特切割位点位于JCV扩增子中,通过切割来区分BKV和JCV。
对17例患有出血性膀胱炎的骨髓移植受者和1例肝移植患者的23份尿液标本进行了BKV和JCV DNA检测。还对4例脑组织标本(石蜡包埋脑组织和1份新鲜冷冻脑活检标本)以及来自3例艾滋病患者和1例肝移植患者的5份脑脊液进行了PCR检测,所有患者均患有进行性多灶性白质脑病(PML)。
该PCR对每种病毒的敏感性为10个基因组。在12例骨髓移植患者的15份尿液标本中检测到BKV DNA。在患有PML的患者的4份脑脊液和4份脑组织中检测到JCV DNA。
我们的结果表明,巢式PCR是一种灵敏且快速的检测方法,可用于诊断BKV和JCV感染。