作者
C Lars Mouritsen, Carl T Wittwer, Christine M Litwin, Liming Yang, Janis J Weis, Thomas B Martins, Troy D Jaskowski, Harry R Hill
发表日期
1996/5/1
期刊
American journal of clinical pathology
卷号
105
期号
5
页码范围
647-654
出版商
Oxford University Press
简介
The authors have developed a simple, nested polymerase chain reaction (PCR) assay for amplification of an outer surface protein A (OspA) gene fragment of Borrelia burgdorferi using rapid temperature cycling and ethidium bromide detection on agarose gels, and applied it to the diagnosis of Lyme disease in humans. With denaturing and annealing temperature spikes instead of holds, cycle times were less than 20 minutes for a 30-cycle amplification. Using this rapid cycle PCR technique, as few as 5 spirochetes per mL of phosphate buffered saline were detected. In addition, B burgdorferi DNA was detected from spirochetes that had been spiked into one of several types of human body fluids including serum, synovial fluid, and cerebrospinal fluid (CSF). A number of clinical samples, which had been tested for Lyme immunoglobulin M (IgM) and immunoglobulin G (IgG) antibody were also examined. In 29 …
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