Gilbert C, Handfield J, Toma E, Lalonde R, Bergeron M G, Boivin G
Department of Microbiology and Research Center in Infectious Diseases, Centre Hospitalier de l'Université Laval, Québec City, Canada.
AIDS. 1998 Jan 22;12(2):125-9. doi: 10.1097/00002030-199802000-00001.
To evaluate the prevalence of the most common cytomegalovirus (CMV) UL97 mutations associated with ganciclovir resistance directly in polymorphonuclear leukocytes (PMNL) of patients with AIDS and CMV retinitis. Also to correlate the presence (or absence) of these mutations with the systemic CMV viral load and the ophthalmologic outcome of these subjects.
Monthly blood samples were obtained from 19 patients with AIDS and CMV retinitis who had been treated with systemic ganciclovir for > or = 2 months. Detection of CMV UL97 mutations was done using nested PCR amplification followed by restriction enzyme analysis. The viral load was assessed with a polymerase chain reaction-based assay and non-isotopic hybridization detection.
CMV UL97 mutations were detected in PMNL of four of 13 (30.8%) patients who had been treated with ganciclovir for > or = 3 months but in none of six patients who had been treated for < 3 months. All four patients with detectable UL97 mutations were presenting evidence of retinitis progression at the time those mutations were first detected (mean, 145.7 days of ganciclovir) and three of four patients had a viral DNA load > 10000 copies per 10(5) PMNL contrasting with the copy numbers in the 15 subjects without mutations (mean, 492.9 copies per 10(5) PMNL after a mean of 146.8 days of ganciclovir).
The prevalence of the most common CMV UL97 mutations associated with ganciclovir resistance in PMNL of patients with AIDS treated for > or = 3 months (30.8%) appears to be higher than the rate of emergence of ganciclovir-resistant CMV isolates as previously reported using phenotypic assays (about 8%). Moreover, the detection of these mutations is associated with a considerable increase in the CMV DNA load in the blood as well as with progression of CMV retinitis during ganciclovir therapy.
直接评估艾滋病合并巨细胞病毒（CMV）视网膜炎患者多形核白细胞（PMNL）中与更昔洛韦耐药相关的最常见CMV UL97突变的发生率。同时将这些突变的存在（或不存在）与全身CMV病毒载量以及这些受试者的眼科结局相关联。
从19例接受全身更昔洛韦治疗≥2个月的艾滋病合并CMV视网膜炎患者中每月采集血样。使用巢式PCR扩增随后进行限制性酶切分析来检测CMV UL97突变。通过基于聚合酶链反应的检测和非同位素杂交检测来评估病毒载量。
在接受更昔洛韦治疗≥3个月的13例患者中的4例（30.8%）的PMNL中检测到CMV UL97突变，但在接受治疗<3个月的6例患者中均未检测到。所有4例可检测到UL97突变的患者在首次检测到这些突变时均有视网膜炎进展的证据（平均更昔洛韦治疗145.7天），4例患者中有3例病毒DNA载量>每10(5)个PMNL 10000拷贝，这与15例无突变受试者的拷贝数形成对比（平均更昔洛韦治疗146.8天后每10(5)个PMNL平均492.9拷贝）。
在接受治疗≥3个月的艾滋病患者的PMNL中，与更昔洛韦耐药相关的最常见CMV UL97突变的发生率（30.8%）似乎高于先前使用表型分析报道的更昔洛韦耐药CMV分离株的出现率（约8%）。此外，这些突变的检测与血液中CMV DNA载量的显著增加以及更昔洛韦治疗期间CMV视网膜炎的进展相关。
