MacGregor R R, Dreyer K, Herman S, Hocknell P K, Nghiem L, Tevere V J, Williams A L
Infectious Diseases Division, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania. 19104-6073, USA.
J Clin Microbiol. 1999 Jan;37(1):90-4. doi: 10.1128/JCM.37.1.90-94.1999.
We evaluated the sensitivity and specificity of a PCR-based qualitative test for the rapid diagnosis of Mycobacterium avium-M. intracellulare complex (MAC) bacteremia in patients with AIDS disease. Eleven subjects with newly culture-proven MAC bacteremia had the following tests performed at biweekly intervals during the first 8 weeks of therapy: blood culture, Mycobacterium-specific PCR, and quantitative human immunodeficiency virus (HIV) viral-load testing. Mycobacterium genus-specific biotinylated primers were used to amplify a sequence of approximately 582 nucleotides within the 16S rRNA genes of M. avium and M. intracellulare. Detection of the amplified product was performed with an oligonucleotide probe-coated microwell plate combined with an avidin-horseradish peroxidase-tetramethylbenzidine conjugate-substrate system. While not as sensitive as BACTEC culture, PCR detected 17 of 18 specimens which grew >/=40 organisms/ml (94.4% sensitivity) and 9 of 16 specimens which grew </=40 organisms/ml (56.3% sensitivity). No clear change in HIV viremia occurred in response to successful treatment of patients' MAC bacteremia. Use of the PCR test allowed detection of MAC bacteremia in 1 day, with a sensitivity similar to those of quantitative blood culture techniques, and it may prove useful for rapid screening of suspected cases. HIV viremia was unaffected by 8 weeks of MAC therapy.
我们评估了一种基于聚合酶链反应(PCR)的定性检测方法在艾滋病患者中快速诊断鸟分枝杆菌-胞内分枝杆菌复合体(MAC)菌血症的敏感性和特异性。11例新确诊为MAC菌血症的患者在治疗的前8周每两周进行一次以下检测:血培养、分枝杆菌特异性PCR以及定量人类免疫缺陷病毒(HIV)病毒载量检测。使用分枝杆菌属特异性生物素化引物扩增鸟分枝杆菌和胞内分枝杆菌16S rRNA基因内约582个核苷酸的序列。通过结合抗生物素蛋白-辣根过氧化物酶-四甲基联苯胺共轭底物系统的寡核苷酸探针包被微孔板检测扩增产物。虽然PCR不如BACTEC培养敏感,但它检测出了18份每毫升生长≥40个菌的标本中的17份(敏感性为94.4%)以及16份每毫升生长≤40个菌的标本中的9份(敏感性为56.3%)。患者的MAC菌血症成功治疗后,HIV病毒血症没有明显变化。使用PCR检测可在1天内检测出MAC菌血症,其敏感性与定量血培养技术相似,可能对快速筛查疑似病例有用。8周的MAC治疗对HIV病毒血症没有影响。