Abstract

HERNANDEZ, Yenney; LOBO, Evelyn; MARTINEZ, Siomara  and  ZAMORA, Loidy. EVALUATION OF DIFFERENT DNA EXTRACTION METHODS FROM MYCOPLASMA TO BE EMPLOYEES IN THE DIAGNOSTIC BY PCR. Rev Salud Anim. [online]. 2009, vol.31, n.2, pp.108-114. ISSN 0253-570X.

The aim of this work was to evaluate different DNA extraction methods of mycoplasma with the use of PCR. The characterization of Mycoplasma arginini strain use as positive control in this test was carried out, by means of morphological studies and biochemical and enzymatic tests, in order to obtain a homogenous culture, which allowed evaluating three mycoplasma DNA extraction methods for their use in the PCR test. Methods based on centrifuge and thermal shock with centrifuge were proven for the DNA extraction in cell culture clinical samples, sera and biopharmaceutical products, which had been tested by culture method and Nucleic Acid Hybridization (NAH). Mycoplasma arginini, strain was characterized and evaluated. Either phenol-chloroform extraction as well as two more simple methods describe above, brought the same results when the PCR assay was carried out using M. arginini strain. It has been demonstrated that the methods based on centrifuge and thermal shock with centrifuge are efficient to obtain Mycoplasma DNA from clinical samples with the use of PCR, comparing these results with those obtained by culture method and NAH.

Keywords : mycoplasmas; diagnostic; cell cultures; PCR.

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