Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, 6500 HB Nijmegen, Netherlands.
BMJ. 2010 Feb 25;340:c1018. doi: 10.1136/bmj.c1018.
The presence of the retrovirus xenotropic murine leukaemia virus-related virus (XMRV) has been reported in peripheral blood mononuclear cells of patients with chronic fatigue syndrome. Considering the potentially great medical and social relevance of such a discovery, we investigated whether this finding could be confirmed in an independent European cohort of patients with chronic fatigue syndrome.
Analysis of a well defined cohort of patients and matched neighbourhood controls by polymerase chain reaction.
Certified (ISO 15189) laboratory of clinical virology in a university hospital in the Netherlands. Population Between December 1991 and April 1992, peripheral blood mononuclear cells were isolated from 76 patients and 69 matched neighbourhood controls. In this study we tested cells from 32 patients and 43 controls from whom original cryopreserved phials were still available.
Detection of XMRV in peripheral blood mononuclear cells by real time polymerase chain reaction assay targeting the XMRV integrase gene and/or a nested polymerase chain reaction assay targeting the XMRV gag gene.
We detected no XMRV sequences in any of the patients or controls in either of the assays, in which relevant positive and negative isolation controls and polymerase chain reaction controls were included. Spiking experiments showed that we were able to detect at least 10 copies of XMRV sequences per 10(5) peripheral blood mononuclear cells by real time as well as by nested polymerase chain reaction, demonstrating high sensitivity of both assays.
This study failed to show the presence of XMRV in peripheral blood mononuclear cells of patients with chronic fatigue syndrome from a Dutch cohort. These data cast doubt on the claim that XMRV is associated with chronic fatigue syndrome in the majority of patients.
报道称,在慢性疲劳综合征患者的外周血单个核细胞中存在嗜异性鼠白血病病毒相关病毒(XMRV)。考虑到这一发现具有潜在的重要医学和社会意义,我们调查了这一发现是否能在一个独立的欧洲慢性疲劳综合征患者队列中得到证实。
通过聚合酶链反应分析一个明确的患者队列和匹配的邻里对照。
荷兰一所大学医院的认证(ISO 15189)临床病毒学实验室。
1991 年 12 月至 1992 年 4 月间,从 76 名患者和 69 名匹配的邻里对照中分离出外周血单个核细胞。在这项研究中,我们测试了 32 名患者和 43 名对照的细胞,他们的原始冷冻小瓶仍然可用。
通过针对 XMRV 整合酶基因的实时聚合酶链反应(PCR)检测和/或针对 XMRV gag 基因的嵌套 PCR 检测,在外周血单个核细胞中检测 XMRV。
我们在任何一种检测方法中都没有检测到患者或对照者的任何 XMRV 序列,包括相关的阳性和阴性分离对照以及聚合酶链反应对照。加标实验表明,我们能够通过实时和嵌套 PCR 检测到至少 10 拷贝的 XMRV 序列/10^5 个外周血单个核细胞,证明了两种检测方法的高灵敏度。
这项研究未能显示荷兰队列中慢性疲劳综合征患者外周血单个核细胞中存在 XMRV。这些数据对 XMRV 与大多数患者慢性疲劳综合征相关的说法提出了质疑。