Hofman Véronique, Selva Eric, Landraud Luce, Sicard Dominique, Vénissac Nicolas, Castillo Laurent, Kermarec Alain, Mouroux Jérôme, Dellamonica Pierre, Hofman Paul
Equipe INSERM 02-15, IFR 50, Faculté de Médecine, 06107 Nice Cedex 02.
Ann Pathol. 2003 Jun;23(3):206-15.
Mycobacterium tuberculosis infection remains a major health problem throughout the world. In France, tuberculosis is endemic, particularly in the Paris area (Ile-de-France) and in the south (Provence Alpes côte d'Azur) where immigration is greater than in other countries in northern Europe. Culture is the gold standard for diagnosis of tuberculosis and is the only method enabling a study of strain sensitivity to treatment. Histology contributes to diagnosis in most cases by revealing typical necrotizing granulomatous lesions. The diagnosis is then confirmed by the detection of acid-fast bacilli with Ziehl-Neelsen staining. However, the Ziehl-Neelsen stain is not sensitive and does not allow identification of different species. The polymerase chain reaction (PCR) DNA amplification method has been used to detect M. tuberculosis in formalin-fixed paraffin-embedded tissues. The aim of the present study was to investigate the value of this method for the diagnosis of M. tuberculosis infection. The results obtained with PCR assay were compared to those obtained with histological and microbiological methods (direct examination and culture). Sixty-three specimens (mainly lymph node and lung specimens) exhibiting a positive culture for M. tuberculosis were analyzed. Tuberculosis granulomas were noted in 32/63 cases, tuberculoid granulomas in 18/63, pyoepitheloid granuloms in 10/63, and non-specific inflammation in 3/63. Ziehl-Neelsen staining was positive in 11/63 cases. PCR assay on tissue sections was positive for M. tuberculosis in 58/63 cases. Controls of the PCR method (granulomas due to other mycobacterial species, foreign body granulomas, sarcoidosis granulomas) were all negative. This study shows that PCR from deparaffinized sections, 1) greatly increases the sensitivity of diagnosis of tuberculosis, 2) enables the diagnosis of M. tuberculosis infection. However, although this method reduces the time to diagnosis, culture remains the gold standard for identification of the mycobacterium and for determining the sensitivity of the isolated strain to treatment.
结核分枝杆菌感染仍是全球主要的健康问题。在法国，结核病呈地方性流行，尤其是在巴黎地区（法兰西岛）和南部（普罗旺斯-阿尔卑斯-蓝色海岸），这些地区的移民数量多于北欧其他国家。培养是结核病诊断的金标准，也是唯一能够研究菌株对治疗敏感性的方法。在大多数情况下，组织学通过揭示典型的坏死性肉芽肿病变有助于诊断。然后通过齐-尼氏染色检测抗酸杆菌来确诊。然而，齐-尼氏染色不敏感，且无法鉴别不同菌种。聚合酶链反应（PCR）DNA扩增方法已用于检测福尔马林固定石蜡包埋组织中的结核分枝杆菌。本研究的目的是探讨该方法在诊断结核分枝杆菌感染中的价值。将PCR检测结果与组织学和微生物学方法（直接检查和培养）的结果进行比较。对63份结核分枝杆菌培养阳性的标本（主要是淋巴结和肺标本）进行了分析。63例中有32例发现结核肉芽肿，18例为结核样肉芽肿，10例为脓性上皮样肉芽肿，3例为非特异性炎症。63例中有11例齐-尼氏染色呈阳性。组织切片的PCR检测在63例中有58例结核分枝杆菌呈阳性。PCR方法的对照（其他分枝杆菌属引起的肉芽肿、异物肉芽肿、结节病肉芽肿）均为阴性。本研究表明，从脱石蜡切片进行PCR检测，1）大大提高了结核病诊断的敏感性，2）能够诊断结核分枝杆菌感染。然而，尽管该方法缩短了诊断时间，但培养仍是鉴定分枝杆菌和确定分离菌株对治疗敏感性的金标准。
