Sanderson J D, Moss M T, Tizard M L, Hermon-Taylor J
Department of Surgery, St George's Hospital Medical School, London.
Gut. 1992 Jul;33(7):890-6. doi: 10.1136/gut.33.7.890.
Crohn's disease has long been suspected of having a mycobacterial cause. Mycobacterium paratuberculosis is a known cause of chronic enteritis in animals, including primates, but may be very difficult to detect by culture. IS900 is a multicopy genomic DNA insertion element highly specific for M paratuberculosis. A polymerase chain reaction (PCR) based on the 5' region of IS900 and capable of the specific detection of a single M paratuberculosis genome was developed. This was applied to DNA extracts of full thickness samples of intestine removed at surgery from 40 patients with Crohn's disease, 23 patients with ulcerative colitis, and 40 control patients without inflammatory bowel disease. Stringent precautions were taken that excluded contamination artefact. M paratuberculosis was identified in 26 of 40 (65%) Crohn's disease, in 1 of 23 (4.3%) ulcerative colitis, and in 5 of 40 (12.5%) control tissues. Positive samples from Crohn's disease were from both the small intestine and colon, those from control tissues were from the colon those from control tissues were from the colon only. All PCR internal control reactions were negative. The presence of M paratuberculosis in a small proportion of apparently normal colonic samples is consistent with a previously unsuspected alimentary prevalence in humans. The presence in two thirds of Crohn's disease tissues but in less than 5% of ulcerative colitis tissues is consistent with an aetiological role for M paratuberculosis in Crohn's disease.
长期以来,人们一直怀疑克罗恩病由分枝杆菌引起。副结核分枝杆菌是包括灵长类动物在内的动物慢性肠炎的已知病因,但通过培养可能很难检测到。IS900是一种对副结核分枝杆菌具有高度特异性的多拷贝基因组DNA插入元件。基于IS900 5'区域开发了一种聚合酶链反应(PCR),能够特异性检测单个副结核分枝杆菌基因组。将其应用于40例克罗恩病患者、23例溃疡性结肠炎患者以及40例无炎症性肠病的对照患者手术切除的肠全层样本的DNA提取物。采取了严格的预防措施以排除污染假象。在40例克罗恩病患者中有26例(65%)检测到副结核分枝杆菌,23例溃疡性结肠炎患者中有1例(4.3%)检测到,40例对照组织中有5例(12.5%)检测到。克罗恩病的阳性样本来自小肠和结肠,对照组织的阳性样本仅来自结肠。所有PCR内部对照反应均为阴性。在一小部分看似正常的结肠样本中存在副结核分枝杆菌,这与之前未被怀疑的人类消化道感染率一致。在三分之二的克罗恩病组织中存在副结核分枝杆菌,但在不到5%的溃疡性结肠炎组织中存在,这与副结核分枝杆菌在克罗恩病病因学中的作用一致。