Aslanzadeh J, de la Viuda M, Fille M, Smith W B, Namdari H
Department of Laboratory Medicine, University of Connecticut Health Center, Farmington 06030, USA.
Mol Cell Probes. 1998 Aug;12(4):207-11. doi: 10.1006/mcpr.1998.0174.
The major drawback in effective use of polymerase chain reaction (PCR) for detecting Mycobacterium tuberculosis (MTB) in clinical samples is the presence of PCR inhibitors and unique cell components of the organism that complicate DNA extraction and subsequent PCR amplification. A PCR assay with a unique multistep DNA extraction method that minimizes these problems was compared in a prospective study to acid-fast bacilli stain (AFBS) and culture for detecting MTB in clinical samples. A total of 254 clinical specimens in two separate studies were processed for MTB by these techniques. While PCR and culture were 100% sensitive and specific, culture required up to 8 weeks of incubation and additional time to perform biochemical testing to identify the isolated micro-organism. Acid-fast bacilli stain had a specificity of about 87% and did not differentiate among Mycobacterial species. In contrast, the results from PCR were available within 48 h and did not require additional testing to attain a final result. Polymerase chain reaction was highly reliable for detection and confirmation and interpretation of positive AFBS results. The assay was easy to perform with a turn around time of about 2 days.
在临床样本中有效使用聚合酶链反应(PCR)检测结核分枝杆菌(MTB)的主要缺点是存在PCR抑制剂以及该生物体独特的细胞成分,这会使DNA提取及后续的PCR扩增变得复杂。在一项前瞻性研究中,将一种采用独特的多步骤DNA提取方法以尽量减少这些问题的PCR检测方法,与抗酸杆菌染色(AFBS)和培养法进行比较,用于检测临床样本中的MTB。在两项独立研究中,共采用这些技术对254份临床标本进行了MTB检测。虽然PCR和培养法的敏感性和特异性均为100%,但培养法需要长达8周的孵育时间以及额外时间来进行生化检测以鉴定分离出的微生物。抗酸杆菌染色的特异性约为87%,且无法区分分枝杆菌种类。相比之下,PCR结果在48小时内即可获得,且无需额外检测就能得出最终结果。聚合酶链反应对于检测、确认及解读抗酸杆菌染色阳性结果高度可靠。该检测方法易于操作,周转时间约为2天。