The polymerase chain reaction forMycoplasma gallisepticumdetection

Abstract

On the basis of the aligned 16S rRNA sequences of Mollicutes, a pair of primers was chosen for the detection of Mycoplasma gallisepticum. When used in the polymerase chain reaction (PCR), the primers detected a specific amplification of all Mg strains tested, yielding an expected 330 bp product. Amplification was not detected when other Mollicutes or E. coli were used as PCR templates. SPF chickens were experimentally inoculated with two strains of M. gallisepticum or Mycoplasma iowae. Tracheal swabs were collected 8, 15, 20 and 28 days after inoculation, and cultured for mycoplasma or tested by PCR. PCR products were detected by hybridization with a digoxigenin-labeled probe and by chemiluminescence. The results showed that culture was positive for 49/73 swabs while PCR detected 70/72 positive samples. Thus, PCR can provide the basis of a sensitive, specific, rapid and non-radio-active method for detecting M. gallisepticum.

Polymerase chain reaction for detection of Mycoplasma gallisepticum in environmental samples

“…PCR amplification of M. gallisepticum DNA was performed as previously described by Kempf et al (1993). Briefly, M. gallisepticum DNA was amplified in 50 ml PCR mixture containing PCR buffer (10 mM Tris-HCl, 50 mM KCl, 2.5 mM MgCl 2 ; pH 8.3), a 500 mM concentration of each deoxyribonucleoside triphosphate (Pharmacia Biotech, Orsay, France), 5 mM tetramethyl ammonium chloride (Sigma Aldrich, Saint Quentin Fallavier, France), 800 nM each primer, 1 U Taq DNA polymerase (Roche-Boehringer, Meylan, France), and 5 ml DNA template.…”

Characterization of Mutations in DNA Gyrase and Topoisomerase IV Involved in Quinolone Resistance of Mycoplasma gallisepticum Mutants Obtained In Vitro

“…Two strains of M. gallisepticum, ATCC 15302, a reference strain (26), and 41-91, a field strain (15), were used in the selection. Strains were grown in Frey agar or broth medium (15).…”

“…Strains were grown in Frey agar or broth medium (15). Flumequine, norfloxacin, ofloxacin, and oxolinic acid used for the MIC determinations were purchased from SigmaAldrich (Saint Quentin Fallavier, France), while danofloxacin came from Pfizer (Amboise, France), marbofloxacin came from Vétoquinol (Magny-Vernois, France), difloxacin came from Fort Dodge Animal Health (Princeton, N.J.), and enrofloxacin and ciprofloxacin came from Bayer Pharma (Puteaux, France).…”

Development of a loop-mediated isothermal amplification targeting a gene within the pyruvate dehydrogenase complex, the pdhA gene, for rapid detection of Mycoplasma gallisepticum

“…12 This PCR method, termed the MG PCR, was compared, in terms of sensitivity and specificity, with the LAMP assay. The primers for the MG PCR assay are shown in Table 2.…”

“…1,6,8 Polymerase chain reaction (PCR) assays, which are characterized by high sensitivity and specificity, are increasingly used in disease diagnosis, with many PCR methods having been established to detect M. gallisepticum. 9,12,24 Real-time PCR methods for the diagnosis of M. gallisepticum have been described. 17,25 However, PCR-based methods have practical problems such as a high capital cost for the instrumentation, 584155V DIXXX10.1177/1040638715584155LAMP for rapid detection of Mycoplasma gallisepticumZhang et al…”