Evaluation of Different PCR Systems for the Detection of Mycoplasma gallisepticum in Chicken Trachea
Serpil Kâhya Demirbilek
Abstract
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Serpil Kâhya Demirbilek
Abstract
Open-access reader
p.p1 {margin: 0.0px 0.0px 0.0px 0.0px; font: 12.0px 'Times New Roman'; min-height: 15.0px} p.p2 {margin: 0.0px 0.0px 4.0px 19.0px; text-align: justify; line-height: 9.1px; font: 9.0px 'Times New Roman'} span.s1 {font: 12.0px 'Times New Roman'} In this work, we detected the MG-serologic condition by rapid plate agglutination tests, used Air Thermal Cycler (ATC PCR) (Idaho Technologies) and LightCycler real-time PCR system (LC PCR) (Roche Diagnostics, Manheim, Germany) for rapid and reliable detection of Mycoplasma gallisepticum (MG) from tracheal swab samples of naturally infected breeder chickens, and determinated MG-DNA detection limit by MG LC PCR from both pure culture and artificially spiked samples. One hundred and seventy seven tracheal swab samples from 16 flocks of 3 different companies were tested by LC PCR. Despite 117 chickens from 10 flocks were diagnosed as MG-seropositive, only 41 (35%) of tracheal swab samples from 3 (%30) flocks were found positive by LC PCR. Sixty (33.8%) of the samples from 6 MG-seronegative flocks were also found to be MG negative by LC PCR. Two hundred twelve MG-seropositive samples from 4 companies (3 of them are same companies tested previously by LC) were tested by ATC PCR and detected only 4 (1.8%) tracheal swab samples MG-positive. The LC PCR gives the results in approximately 6 hours DNA extraction, and is rapid and reliable confirmation and detection test ready to be implemented for screening MG-infected flocks in poultry companies.
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p.p1 {margin: 0.0px 0.0px 0.0px 0.0px; font: 12.0px 'Times New Roman'; min-height: 15.0px} p.p2 {margin: 0.0px 0.0px 4.0px 19.0px; text-align: justify; line-height: 9.1px; font: 9.0px 'Times New Roman'} span.s1 {font: 12.0px 'Times New Roman'} In this work, we detected the MG-serologic condition by rapid plate agglutination tests, used Air Thermal Cycler (ATC PCR) (Idaho Technologies) and LightCycler real-time PCR system (LC PCR) (Roche Diagnostics, Manheim, Germany) for rapid and reliable detection of Mycoplasma gallisepticum (MG) from tracheal swab samples of naturally infected breeder chickens, and determinated MG-DNA detection limit by MG LC PCR from both pure culture and artificially spiked samples. One hundred and seventy seven tracheal swab samples from 16 flocks of 3 different companies were tested by LC PCR. Despite 117 chickens from 10 flocks were diagnosed as MG-seropositive, only 41 (35%) of tracheal swab samples from 3 (%30) flocks were found positive by LC PCR. Sixty (33.8%) of the samples from 6 MG-seronegative flocks were also found to be MG negative by LC PCR. Two hundred twelve MG-seropositive samples from 4 companies (3 of them are same companies tested previously by LC) were tested by ATC PCR and detected only 4 (1.8%) tracheal swab samples MG-positive. The LC PCR gives the results in approximately 6 hours DNA extraction, and is rapid and reliable confirmation and detection test ready to be implemented for screening MG-infected flocks in poultry companies.
Key concepts: Flock, Mycoplasma gallisepticum, Biology, Veterinary medicine, Serology, Polymerase chain reaction, Detection limit, DNA extraction