Oberhelman Richard A, Soto-Castellares Giselle, Caviedes Luz, Castillo Maria E, Kissinger Patricia, Moore David A J, Evans Carlton, Gilman Robert H
Department of Tropical Medicine, Tulane School of Public Health and Tropical Medicine, 1440 Canal St, SL 29, New Orleans, Louisiana 70112, USA.
Pediatrics. 2006 Jul;118(1):e100-6. doi: 10.1542/peds.2005-2623. Epub 2006 Jun 2.
The diagnosis of pulmonary tuberculosis presents challenges in children, because symptoms are nonspecific, sputa are not accessible, and Mycobacterium tuberculosis cultures and smears often are negative. The Microscopic Observation Drug Susceptibility technique is a simple, inexpensive method for Mycobacterium tuberculosis isolation with superior speed and sensitivity over Lowenstein-Jensen culture in studies of adults with pulmonary tuberculosis. The objective of this study was to determine whether Microscopic Observation Drug Susceptibility culture can improve the sensitivity and the speed of Mycobacterium tuberculosis recovery among Peruvian children with symptoms suggestive of pulmonary tuberculosis.
Two specimens of each type (gastric aspirate, nasopharyngeal aspirate, and stool specimens) were collected from each patient, examined by auramine stain, and cultured by Microscopic Observation Drug Susceptibility and Lowenstein-Jensen techniques. Patients (n = 165) were enrolled between April 2002 and February 2004 at the Instituto de Salud del Niño, the major pediatric hospital in Lima, Peru. Inclusion criteria were age < or = 12 years, Stegen-Toledo clinical score > or = 5 points, and absence of antituberculous therapy. The main outcome measurements were (1) proportion of specimens that were culture positive by Microscopic Observation Drug Susceptibility versus Lowenstein-Jensen and (2) days required for positive culture result, stratified by specimen type and auramine stain result.
Fifteen (9%) patients had at least 1 positive Mycobacterium tuberculosis culture (from stool in 3 cases, nasopharyngeal aspirate in 8 cases, and gastric aspirate in 15 cases). Thirty-eight culture-positive specimens were obtained (22 gastric aspirate, 12 nasopharyngeal aspirates, and 4 stools). Microscopic Observation Drug Susceptibility provided significantly more positive cultures than Lowenstein-Jensen (33 of 38 specimens culture positive by Microscopic Observation Drug Susceptibility vs 21 of 38 by Lowenstein-Jensen). This was attributed to enhanced recovery of Mycobacterium tuberculosis from auramine-negative specimens (19 of 23 by Microscopic Observation Drug Susceptibility vs 9 of 23 by Lowenstein-Jensen), in contrast to similar detection rates for the 2 tests with auramine-positive samples. Similar results were found for analyses that were limited to gastric aspirates. Isolation was faster by Microscopic Observation Drug Susceptibility than Lowenstein-Jensen.
Isolation of Mycobacterium tuberculosis from children with suspected pulmonary tuberculosis by Microscopic Observation Drug Susceptibility demonstrated greater yield and faster recovery than by Lowenstein-Jensen method, significantly improving local capabilities to detect pediatric tuberculosis in resource-poor settings.
儿童肺结核的诊断颇具挑战,因为症状不具特异性,难以获取痰液,且结核分枝杆菌培养和涂片结果常为阴性。显微镜观察药物敏感性技术是一种简单、廉价的结核分枝杆菌分离方法,在对成人肺结核的研究中,其速度和敏感性优于罗 - 琴培养法。本研究的目的是确定显微镜观察药物敏感性培养能否提高秘鲁有肺结核疑似症状儿童结核分枝杆菌的检出率及速度。
从每位患者采集两种类型的标本(胃抽吸物、鼻咽抽吸物和粪便标本),进行金胺染色检查,并采用显微镜观察药物敏感性技术和罗 - 琴技术进行培养。2002年4月至2004年2月期间,在秘鲁利马的主要儿科医院——儿童健康研究所招募了165名患者。纳入标准为年龄≤12岁、Stegen - Toledo临床评分≥5分且未接受抗结核治疗。主要观察指标为:(1)显微镜观察药物敏感性培养与罗 - 琴培养的标本培养阳性比例;(2)培养结果呈阳性所需天数,按标本类型和金胺染色结果分层。
15名(9%)患者至少有1次结核分枝杆菌培养阳性(3例来自粪便,8例来自鼻咽抽吸物,15例来自胃抽吸物)。共获得38份培养阳性标本(22份胃抽吸物、12份鼻咽抽吸物和4份粪便)。显微镜观察药物敏感性培养获得的阳性培养物显著多于罗 - 琴培养(显微镜观察药物敏感性培养的38份标本中有33份培养阳性,罗 - 琴培养的38份中有21份)。这归因于从金胺染色阴性标本中结核分枝杆菌的回收率提高(显微镜观察药物敏感性培养的23份中有19份,罗 - 琴培养的23份中有9份),而两种检测方法对金胺染色阳性样本的检出率相似。对仅限于胃抽吸物的分析也得到了类似结果。显微镜观察药物敏感性培养的分离速度比罗 - 琴培养快。
通过显微镜观察药物敏感性技术从疑似肺结核儿童中分离结核分枝杆菌,其产量更高且恢复速度更快,显著提高了资源匮乏地区检测儿童结核病的能力。