Guy E C, Stanek G
Public Health Laboratory, General Hospital, Southampton.
J Clin Pathol. 1991 Jul;44(7):610-1. doi: 10.1136/jcp.44.7.610.
Borrelia burgdorferi, the causative agent of Lyme disease, was detected in patients' serum by DNA amplification using the polymerase chain reaction (PCR). B burgdorferi was pelleted from serum samples by centrifugation (10,000 x g for 10 minutes) and lysed by treatment with ammonium hydroxide (100 degrees C for 15 minutes). Two pairs of "nested" PCR primers complementary to the gene encoding a major outer surface protein (OSP A) of B burgdorferi were used in DNA amplification under standard PCR conditions (Perkin-Elmer Cetus). Two out of five patients with erythema migrans, the characteristic primary skin lesion associated with early Lyme disease, were positive by the PCR. This method could form the basis of a useful routine laboratory test in those cases of early Lyme disease where conventional serological testing commonly yields equivocal or false negative results.
通过聚合酶链反应（PCR）进行DNA扩增，在患者血清中检测到莱姆病的病原体——伯氏疏螺旋体。将血清样本通过离心（10,000×g，10分钟）沉淀出伯氏疏螺旋体，并用氢氧化铵处理（100℃，15分钟）使其裂解。在标准PCR条件下（珀金-埃尔默塞特斯公司），使用两对与编码伯氏疏螺旋体主要外表面蛋白（OSP A）的基因互补的“巢式”PCR引物进行DNA扩增。在五例游走性红斑患者中，有两例检测出该特征性原发性皮肤损害与早期莱姆病相关，PCR检测呈阳性。对于那些常规血清学检测通常产生不确定或假阴性结果的早期莱姆病病例，该方法可作为一项有用的常规实验室检测的基础。
