Chantranuwat Chavit, Assanasen Thamathorn, Shuangshoti Shanop, Sampatanukul Pichet
King Chulalongkorn Memorial Hospital, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Southeast Asian J Trop Med Public Health. 2006 Sep;37(5):940-7.
A polymerase chain reaction (PCR) protocol for detecting IS6110 repetitive insertion sequence of Mycobacterium tuberculosis (MTB) was tested on archival Papanicolaou (Pap)-stained fine needle aspirated (FNA) smears from 24 patients with cervical tuberculous lymphadenopathy and 30 negative controls. The protocol involved protease digestion or phenolchloroform extraction, and simple or nested PCR, with PCR amplification of human beta-globin gene for internal control of DNA quality. Sensitivity of 50% and specificity of 100% were obtained. Sensitivity in smears showing necrosis without granuloma was 70% (7/10), whereas it was 36% (5/14) in smears with presence of granuloma. On the other hand, sensitivity of 18% (4/22) was obtained using FNA acid-fast stain, 25% (1/4) for acid-fast stain in histological section, 50% (2/4) for culture, and 100% (8/8) for PCR of fresh specimens. PCR for MTB detection in Papanicolaou-stained slides is a practical and valuable method when no fresh specimen but only Pap-stained smear is available.
一种用于检测结核分枝杆菌(MTB)IS6110重复插入序列的聚合酶链反应(PCR)方案,在24例宫颈结核性淋巴结炎患者的巴氏(Pap)染色细针穿刺抽吸(FNA)涂片存档标本以及30份阴性对照上进行了测试。该方案包括蛋白酶消化或酚氯仿提取,以及简单PCR或巢式PCR,并通过PCR扩增人β-珠蛋白基因来内控DNA质量。获得了50%的灵敏度和100%的特异性。在显示无肉芽肿坏死的涂片中灵敏度为70%(7/10),而在有肉芽肿的涂片中灵敏度为36%(5/14)。另一方面,使用FNA抗酸染色的灵敏度为18%(4/22),组织切片抗酸染色为25%(1/4),培养为50%(2/4),新鲜标本PCR为100%(8/8)。当没有新鲜标本而只有巴氏染色涂片可用时,在巴氏染色载玻片上进行MTB检测的PCR是一种实用且有价值的方法。