Matar G M, Khneisser I A, Abdelnoor A M
Department of Microbiology and Immunology, Faculty of Medicine, American University of Beirut, New York 10022, USA.
J Clin Microbiol. 1996 Feb;34(2):477-8. doi: 10.1128/jcm.34.2.477-478.1996.
We developed a PCR-based assay for the rapid and specific laboratory diagnosis of human brucellosis directly from whole blood. Specimens were collected in EDTA tubes from 17 patients with acute serologic brucellosis and 3 patients with chronic relapsing brucellosis as determined by serologic tests and the patient's clinical picture. DNA was extracted from peripheral mononuclear cells obtained from the blood of patients with brucellosis and control individuals. Specific primers for the PCR amplification of a 223-bp region on the sequence encoding the 31-kDa immunogenic Brucella abortus protein (BCSP 31) were used. All amplicons had the expected size of 223 bp. The specificity of amplification was determined by Southern hybridization and restriction endonuclease analysis. DNA extracted from blood taken from 30 healthy individuals as well as from 9 patients with typhoid fever did not show any amplification with the primers used. The test proved to be rapid and specific for the laboratory confirmation of acute human brucellosis. Further studies must be conducted to assess the utility of this test on additional patients with chronic relapsing brucellosis as well as patients under treatment.
我们开发了一种基于聚合酶链反应（PCR）的检测方法，可直接从全血中快速、特异性地对人类布鲁氏菌病进行实验室诊断。通过血清学检测和患者临床表现确诊，从17例急性血清学布鲁氏菌病患者和3例慢性复发性布鲁氏菌病患者的乙二胺四乙酸（EDTA）管中采集标本。从布鲁氏菌病患者和对照个体血液中获取的外周单核细胞中提取DNA。使用特异性引物对编码31 kDa免疫原性流产布鲁氏菌蛋白（BCSP 31）的序列上的一个223 bp区域进行PCR扩增。所有扩增产物大小均为预期的223 bp。通过Southern杂交和限制性内切酶分析确定扩增的特异性。从30名健康个体以及9名伤寒患者采集的血液中提取的DNA，使用所用引物未显示任何扩增。该检测方法被证明对急性人类布鲁氏菌病的实验室确诊快速且特异。必须进行进一步研究，以评估该检测方法对其他慢性复发性布鲁氏菌病患者以及正在接受治疗患者的实用性。
