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Journal of Clinical Microbiology, September 1998, p. 2443-2446, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Strategy for Optimizing DNA Amplification in a Peripheral Blood PCR Assay Used for Diagnosis of Human Brucellosis

Pilar Morata,1,* Maria Isabel Queipo-Ortuño,1 and Juan de Dios Colmenero2

Department of Biochemistry and Molecular Biology, Málaga University,1 and Infectious Diseases Unit, "Carlos Haya" Regional Hospital,2 Málaga, Spain

Received 2 January 1998/Returned for modification 10 April 1998/Accepted 11 June 1998

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 µg, thereby avoiding false-negative results.

* Corresponding author. Mailing address: Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Málaga, 29071 Málaga, Spain. Phone: 34 95 2131538. Fax: 34 95 2131534. E-mail: Morata{at}uma.es.

Journal of Clinical Microbiology, September 1998, p. 2443-2446, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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