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鸟分枝杆菌复合群的聚合酶链反应-杂交检测法：人类外周血检测的优化

PCR-hybridization assay for Mycobacterium avium complex: optimization of detection in peripheral blood from humans.

作者信息

Ferrario G, Gori A, Rossi A, Catozzi L, Molteni C, Marchetti G, Bandera A, Rossi M C, Esposti A D, Franzetti F

机构信息

Institute of Infectious Diseases and Tropical Medicine, Luigi Sacco Hospital, University of Milan, Milan, Varese, Italy.

出版信息

J Clin Microbiol. 2001 Apr;39(4):1638-43. doi: 10.1128/JCM.39.4.1638-1643.2001.

DOI:10.1128/JCM.39.4.1638-1643.2001

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC87988/

Abstract

We evaluated the sensitivity of a DNA amplification test for the detection of Mycobacterium avium in blood samples using different blood components and different DNA extraction methods. M. avium-inoculated blood samples were processed to obtain separate blood components: peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNCs), and whole-blood sodium dodecyl sulfate (SDS)-lysate pellets. The sensitivity for the detection of the lowest mycobacterial load (1 CFU/ml) was significantly greater (P < 0.01) with DNA extracted from SDS-lysate pellets than with DNA extracted from PBMCs or PMNCs. Subsequently, DNA extraction methods based on guanidine NaOH, and proteinase were compared. The sensitivity of the guanidine-based method was significantly greater (P < 0.01) than those of the others.

摘要

我们使用不同的血液成分和不同的DNA提取方法，评估了一种DNA扩增试验检测血液样本中鸟分枝杆菌的敏感性。对接种了鸟分枝杆菌的血液样本进行处理，以获得不同的血液成分：外周血单核细胞（PBMC）、多形核细胞（PMNC）和全血十二烷基硫酸钠（SDS）裂解物沉淀。从SDS裂解物沉淀中提取的DNA检测最低分枝杆菌载量（1 CFU/ml）的敏感性显著高于从PBMC或PMNC中提取的DNA（P < 0.01）。随后，比较了基于氢氧化钠胍和蛋白酶的DNA提取方法。基于胍的方法的敏感性显著高于其他方法（P < 0.01）。