白细胞浓度在巨细胞病毒(CMV)抗原血症定量检测中的意义

Steven M Lipson , Ana Toro , Madhavi Lotlikar , Mark E Match , Mark H Kaplan , David H Shepp , Jerry Gong
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引用次数: 8

摘要

背景:巨细胞病毒抗原血症(CMV-Ag)定量检测是获得性免疫缺陷综合征(艾滋病)和其他免疫功能低下患者群体护理和管理中使用的一项重要技术。这种测定的性能取决于对外周血的多形核白细胞(PMNL)室的适当处理。然而,CMV-Ag检测性能的细胞输入标准尚未建立。使用CMV-Ag测定的实验室之间的解释差异可能反映了这种测试一致性的缺乏。目的:探讨不同浓度PMNL对外周血巨细胞病毒(CMV)定量的影响。产生最佳病毒检出率的白细胞浓度将与壳瓶测定法-间接免疫荧光测定法(SVA-IFA)和常规管培养法(TC-CPE)进行比较。研究设计:对74份新鲜采集的血液标本进行CMV-Ag检测,使用细胞自旋制剂2×105, 4×105和在初步实验中,8×105 PMNLs/slide。将这些研究获得的数据与SVA-IFA和TC-CPE进行比较。通过在更昔洛韦(GCV)、膦酸钠(PFA)或联合药物治疗开始时对11例有症状患者进行序贯检测,监测病毒载量。结果:在74份CMV-Ag检测的血液标本中,含有4×105的细胞自旋制剂与2×105 PMNLs/slide相比,平均阳性细胞增加215% (P=0.03)。含有8×105细胞的PMNL载玻片制备产生的背景水平妨碍了载玻片的准确读取。CMV-Ag检测灵敏度高于SVA-IFA,但与TC-CPE相当。在开始药物治疗的11例患者中,有8例在2-3周内病毒载量明显降低;3例患者(2例在治疗后3周内死亡)病毒载量没有下降。一名患者被鉴定为携带PFA耐药菌株。结论:4×105细胞的PMNL浓度有助于CMV-Ag检测载玻片制备的读取。此外,改进的CMV- ag检测方法可用于监测病毒载量,以跟踪敏感或耐药的CMV毒株。
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Significance of leukocyte concentration in the performance of the quantitative cytomegalovirus (CMV) antigenemia assay

Background: The quantitative cytomegalovirus-antigenemia (CMV-Ag) assay is an important technology in the regimen of tests utilized in the care and management of acquired immunodeficiency syndrome (AIDS) and other immunocompromised patient groups. Performance of this assay is contingent upon the appropriate processing of the polymorphonuclear leukocyte (PMNL) compartment of the peripheral blood. However, a cell input standard in the performance of the CMV-Ag assay, has not been established. Interpretive differences between laboratories utilizing the CMV-Ag assay may reflect this lack of test uniformity.

Objectives: To determine the effect of different PMNL concentrations on the quantitation of CMV in peripheral blood. The leukocyte concentration resulting in optimal rates of viral detection, will be compared with the shell vial assay-indirect immunofluorescent assay (SVA-IFA), and conventional tube culture (TC-CPE).

Study design: A total of 74 freshly collected blood specimens were tested by the CMV-Ag assay, using cytospin preparations consisting of 2×105, 4×105 and in preliminary experiments, 8×105 PMNLs/slide. Data obtained from these studies were compared to SVA-IFA and TC-CPE. Viral load was monitored among 11 symptomatic patients through sequential testing of these patients at the start of ganciclovir (GCV), foscarnet (PFA), or combination drug therapy.

Results: Among 74 blood specimens tested by the CMV-Ag assay, cytospin preparations consisting of 4×105 compared with 2×105 PMNLs/slide, affected a mean positive cell increase of 215% (P=0.03). PMNL slide preparations consisting 8×105 cells produced background levels which prevented accurate reading of slides. The CMV-Ag assay was more sensitive than the SVA-IFA, but equivalent to TC-CPE. Among 11 patients started on drug therapy, viral load was markedly reduced in 8 within 2–3 weeks; three patients (2 deceased within 3 weeks after receiving therapy), showed no decrease in viral load. One patient was identified as harboring a PFA resistant strain.

Conclusions: A PMNL concentration of 4×105 cells facilitated the reading of CMV-Ag assay slide preparations. The modified CMV-Ag assay furthermore, is applicable in the monitoring of viral load for the tracking of susceptible or resistant CMV strains.

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