Scarpellini P, Racca S, Cinque P, Delfanti F, Gianotti N, Terreni M R, Vago L, Lazzarin A
Infectious Diseases Department, San Raffaele Scientific Institute, Milan, Italy.
AIDS. 1995 Aug;9(8):895-900. doi: 10.1097/00002030-199508000-00010.
To investigate the usefulness of polymerase chain reaction (PCR) from cerebrospinal fluid (CSF) for rapid diagnosis and assessing treatment response of tuberculous meningitis (TBM) in AIDS patients.
Forty-four CSF samples from 10 patients with TBM confirmed by autopsy or by a culture of CSF (41 samples) and from two patients with highly probable TBM (three samples) were analysed. CSF specimens were collected before and during standard antituberculous treatment. CSF samples from 24 AIDS patients with autopsy evidence of other neurologic diseases were studied as controls.
A nested PCR amplifying a 123 base-pair fragment of the IS6110 sequence was developed. Heating to 95 degrees C for 15 min was used for pre-PCR treatment of samples.
Detection limit was 10(2) colony-forming units per ml or 10 fg purified Mycobacterium tuberculosis DNA. M.tuberculosis DNA was detected in CSF from all the 12 confirmed or highly probable TBM cases. CSF was positive by nested PCR in 17 of 17 (100%) and 18 of 27 (67%) samples collected before and during therapy, respectively. Clinical and microbiological follow-up > or = 2 weeks was available for seven patients. PCR-positive CSF converted to M. tuberculosis DNA negative in four patients that showed improvement during treatment, but it remained positive in three patients who died of disseminated tuberculosis. All the CSF samples from the non-TBM controls were negative by nested PCR.
Nested PCR for detection of M. tuberculosis DNA is specific for diagnosis of TBM and more sensitive than conventional bacteriology. Moreover, nested PCR could be a useful method for assessing treatment response in AIDS patients with TBM.
探讨脑脊液聚合酶链反应（PCR）在艾滋病患者结核性脑膜炎（TBM）快速诊断及评估治疗反应中的作用。
对10例经尸检或脑脊液培养确诊为TBM患者的44份脑脊液样本（41份样本）以及2例高度疑似TBM患者的3份样本进行分析。脑脊液样本在标准抗结核治疗前及治疗期间采集。选取24例有其他神经系统疾病尸检证据的艾滋病患者的脑脊液样本作为对照。
建立一种巢式PCR方法，扩增IS6110序列的123个碱基对片段。样本进行PCR前处理时加热至95℃ 15分钟。
检测限为每毫升10²个菌落形成单位或10 fg纯化的结核分枝杆菌DNA。12例确诊或高度疑似TBM病例的脑脊液中均检测到结核分枝杆菌DNA。治疗前和治疗期间采集的脑脊液样本，巢式PCR检测阳性率分别为17/17（100%）和27/18（67%）。7例患者有≥2周的临床和微生物学随访。4例治疗期间病情改善的患者，PCR阳性的脑脊液转为结核分枝杆菌DNA阴性，但3例死于播散性结核的患者脑脊液仍为阳性。所有非TBM对照的脑脊液样本巢式PCR检测均为阴性。
巢式PCR检测结核分枝杆菌DNA对TBM诊断具有特异性，且比传统细菌学方法更敏感。此外，巢式PCR可作为评估艾滋病合并TBM患者治疗反应的有用方法。
