Lengerova Martina, Racil Zdenek, Volfova Pavlina, Lochmanova Jana, Berkovcova Jitka, Dvorakova Dana, Vorlicek Jiri, Mayer Jiri
Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematooncology, University Hospital Brno, Brno, Czech Republic.
J Clin Microbiol. 2007 Mar;45(3):1042-4. doi: 10.1128/JCM.01109-06. Epub 2007 Jan 17.
Here we report how variability in the human cytomegalovirus genome sequence may seriously affect the outcome of its molecular diagnosis. A real-time quantitative PCR assay targeting the major immediate-early gene failed to detect the viral load in some, but not all, clinical samples from hematooncological patients. By amplification and sequencing the DNA across the regions targeted by this assay we found a number of nucleotide substitutions which accounted for decreased primer/probe binding. This decreased the sensitivity of the assay up to 1,000-fold.
在此我们报告人类巨细胞病毒基因组序列的变异性如何可能严重影响其分子诊断结果。一种针对主要立即早期基因的实时定量PCR检测方法未能在一些(但并非全部)血液肿瘤患者的临床样本中检测到病毒载量。通过对该检测方法所针对区域的DNA进行扩增和测序,我们发现了一些核苷酸替换,这些替换导致引物/探针结合减少。这使该检测方法的灵敏度降低了高达1000倍。