Boivin G, Chou S, Quirk M R, Erice A, Jordan M C
Department of Microbiology, Université Laval, Quebec City, Canada.
J Infect Dis. 1996 Mar;173(3):523-8. doi: 10.1093/infdis/173.3.523.
Cytomegalovirus (CMV) UL97 mutations associated with ganciclovir resistance at codons 460, 594, and 595 were detected by polymerase chain reaction (PCR) followed by restriction enzyme analysis in CMV blood isolates and directly in polymorphonuclear leukocyte (PMNL) DNA extracts of 4 subjects who died of progressive disseminated CMV disease due to ganciclovir-resistant CMV strains. The CMV DNA load was also serially determined in leukocyte fractions of these patients using a quantitative-competitive PCR assay. There was excellent concordance between specific UL97 mutations in blood culture isolates and those detected in PMNL fractions for all patients. Emergence of such UL97 mutations during ganciclovir therapy was associated with an increasing CMV DNA burden in leukocytes of the 2 patients with AIDS but not in the 2 subjects with chronic lymphocytic leukemia. Rapid molecular strategies, including detection of common CMV UL97 mutations and CMV DNA quantitation, can be used directly in leukocytes of immunocompromised subjects with CMV disease to monitor antiviral therapy.
通过聚合酶链反应（PCR），随后进行限制性酶切分析，在巨细胞病毒（CMV）血液分离株中以及直接在4名因耐更昔洛韦的CMV毒株导致进行性播散性CMV疾病死亡的受试者的多形核白细胞（PMNL）DNA提取物中，检测到了与更昔洛韦耐药相关的CMV UL97在460、594和595密码子处的突变。使用定量竞争PCR检测法，还对这些患者白细胞组分中的CMV DNA载量进行了连续测定。所有患者血液培养分离株中的特定UL97突变与在PMNL组分中检测到的突变之间具有极好的一致性。在更昔洛韦治疗期间出现此类UL97突变，与2例艾滋病患者白细胞中CMV DNA负担增加有关，但与2例慢性淋巴细胞白血病患者无关。包括检测常见CMV UL97突变和CMV DNA定量在内的快速分子策略，可直接用于患有CMV疾病的免疫受损受试者的白细胞中，以监测抗病毒治疗。
