Gerna G, Baldanti F, Sarasini A, Furione M, Percivalle E, Revello M G, Zipeto D, Zella D
Virus Laboratory, University of Pavia, Italy.
Antimicrob Agents Chemother. 1994 Jan;38(1):38-44. doi: 10.1128/AAC.38.1.38.
The aim of this study was to investigate peripheral blood polymorphonuclear leukocytes and, whenever possible, aqueous humor from 65 AIDS patients with ophthalmoscopically diagnosed human cytomegalovirus (HCMV) retinitis to determine (i) whether patients consistently carry viral DNA and (ii) to what extent foscarnet induction treatment decreases viral DNA levels. HCMV DNA was quantified by PCR using densitometric analysis of hybridization products obtained from external standards and a standard curve from which the number of genome equivalents of test samples, normalized by using an internal amplification control, was interpolated. Results showed that 56 of 65 patients (86.1%) were positive for HCMV DNA prior to induction treatment. Of 41 of the 56 patients (73.2%) whose blood had become DNA negative after induction, only 5 had a high viral load (> 5,000 genome equivalents per 2 x 10(5) polymorphonuclear leukocytes) prior to induction, whereas as many as 13 of the 15 (26.8%) patients remaining DNA positive after induction had a high viral load prior to induction. Finally, of the nine patients (13.8%) with DNA-negative blood prior to induction treatment, three were shifted to foscarnet from ganciclovir, while six were erroneously enrolled in the study. Pre- and postinduction aqueous humor samples were obtained from 12 patients; all of these were DNA positive prior to induction, whereas after induction, 4 became negative, 6 showed a marked decrease in viral DNA, and 2 had nearly stable low DNA levels. In conclusion, PCR is a valuable tool for etiologic diagnosis and monitoring of HCMV retinitis treatment in AIDS patients. HCMV DNA is consistently present in the blood and aqueous humor of all patients with HCMV retinitis. Foscarnet induction treatment is highly effective in suppressing or reducing DNA levels in both blood leukocytes and aqueous humor.
本研究旨在调查65例经眼底镜诊断为人类巨细胞病毒(HCMV)视网膜炎的艾滋病患者的外周血多形核白细胞,并尽可能检测房水,以确定:(i)患者是否持续携带病毒DNA;(ii)膦甲酸钠诱导治疗在多大程度上降低病毒DNA水平。通过PCR对HCMV DNA进行定量分析,采用密度分析法分析从外部标准品获得的杂交产物,并根据标准曲线内插经内部扩增对照标准化后的测试样品基因组当量数。结果显示,65例患者中有56例(86.1%)在诱导治疗前HCMV DNA呈阳性。在诱导后血液变为DNA阴性的56例患者中的41例(73.2%)中,只有5例在诱导前病毒载量较高(每2×10⁵多形核白细胞中>5000个基因组当量),而在诱导后仍为DNA阳性的15例患者中的13例(26.8%)在诱导前病毒载量较高。最后,在诱导治疗前血液DNA阴性的9例患者(13.8%)中,3例从更昔洛韦改用膦甲酸钠,而6例错误地纳入了本研究。从12例患者中获取了诱导前后的房水样本;所有这些样本在诱导前均为DNA阳性,而诱导后,4例变为阴性,6例病毒DNA显著下降,2例病毒DNA水平几乎稳定在低水平。总之,PCR是艾滋病患者HCMV视网膜炎病因诊断和治疗监测的有价值工具。所有HCMV视网膜炎患者的血液和房水中均持续存在HCMV DNA。膦甲酸钠诱导治疗在抑制或降低血液白细胞和房水中的DNA水平方面非常有效。