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用于人类布鲁氏菌病诊断的外周血PCR检测中优化DNA扩增的策略。

Strategy for optimizing DNA amplification in a peripheral blood PCR assay used for diagnosis of human brucellosis.

作者信息

Morata P, Queipo-Ortuño M I, de Dios Colmenero J

机构信息

Department of Biochemistry and Molecular Biology, Málaga University, Málaga, Spain.

出版信息

J Clin Microbiol. 1998 Sep;36(9):2443-6. doi: 10.1128/JCM.36.9.2443-2446.1998.

DOI:10.1128/JCM.36.9.2443-2446.1998
PMID:9705371
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC105141/
Abstract

We studied two of the possible factors which can interfere with specific DNA amplification in a peripheral-blood PCR assay used for the diagnosis of human brucellosis. We found that high concentrations of leukocyte DNA and heme compounds inhibit PCR. These inhibitors can be efficiently suppressed by increasing the number of washings to four or five and decreasing the amount of total DNA to 2 to 4 microg, thereby avoiding false-negative results.

摘要

我们研究了在用于诊断人类布鲁氏菌病的外周血聚合酶链反应(PCR)检测中可能干扰特异性DNA扩增的两个因素。我们发现高浓度的白细胞DNA和血红素化合物会抑制PCR。通过将洗涤次数增加到四到五次,并将总DNA量减少到2至4微克,可以有效抑制这些抑制剂,从而避免假阴性结果。

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J Clin Microbiol. 1998 Sep;36(9):2443-6. doi: 10.1128/JCM.36.9.2443-2446.1998.
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本文引用的文献

1
Rapid diagnosis of human brucellosis by peripheral-blood PCR assay.通过外周血PCR检测快速诊断人类布鲁氏菌病
J Clin Microbiol. 1997 Nov;35(11):2927-30. doi: 10.1128/jcm.35.11.2927-2930.1997.
2
Rapid diagnosis of Mycobacterium tuberculosis bacteremia by PCR.通过聚合酶链反应快速诊断结核分枝杆菌菌血症
J Clin Microbiol. 1996 Mar;34(3):512-5. doi: 10.1128/jcm.34.3.512-515.1996.
3
Hydrogen peroxide decomposes the heme compound in forensic specimens and improves the efficiency of PCR.过氧化氢可分解法医样本中的血红素化合物,并提高聚合酶链反应(PCR)的效率。
Biotechniques. 1996 Sep;21(3):392-4. doi: 10.2144/96213bm10.
4
Evaluation of a novel PCR-based diagnostic assay for detection of Mycobacterium tuberculosis in sputum samples.一种基于聚合酶链反应(PCR)的新型诊断检测方法用于痰液样本中结核分枝杆菌检测的评估。
J Clin Microbiol. 1996 Sep;34(9):2307-8. doi: 10.1128/jcm.34.9.2307-2308.1996.
5
Host DNA can interfere with detection of Borrelia burgdorferi in skin biopsy specimens by PCR.宿主DNA可通过聚合酶链反应(PCR)干扰皮肤活检标本中伯氏疏螺旋体的检测。
J Clin Microbiol. 1996 Apr;34(4):980-2. doi: 10.1128/jcm.34.4.980-982.1996.
6
Rapid laboratory confirmation of human brucellosis by PCR analysis of a target sequence on the 31-kilodalton Brucella antigen DNA.通过对31千道尔顿布鲁氏菌抗原DNA上的靶序列进行PCR分析,快速在实验室确诊人类布鲁氏菌病。
J Clin Microbiol. 1996 Feb;34(2):477-8. doi: 10.1128/jcm.34.2.477-478.1996.
7
Identification of the heme compound copurified with deoxyribonucleic acid (DNA) from bloodstains, a major inhibitor of polymerase chain reaction (PCR) amplification.从血迹中与脱氧核糖核酸(DNA)共纯化的血红素化合物的鉴定,该化合物是聚合酶链反应(PCR)扩增的主要抑制剂。
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Purification of highly degraded DNA by gel filtration for PCR.
Biotechniques. 1994 Feb;16(2):235-8.
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Polymerase chain reaction for detection of Borrelia burgdorferi DNA in skin lesions of early and late Lyme borreliosis.用于检测早期和晚期莱姆病螺旋体病皮肤病变中伯氏疏螺旋体DNA的聚合酶链反应
Eur J Clin Microbiol Infect Dis. 1995 Jan;14(1):1-5. doi: 10.1007/BF02112610.
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A simple salting out procedure for extracting DNA from human nucleated cells.一种从人有核细胞中提取DNA的简单盐析方法。
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