Polymerase chain reaction for detection of Mycoplasma gallisepticum.
A species-specific 760-base pair (bp) BamHI to EcoRI DNA fragment (fMG-2) was isolated from a Mycoplasma gallisepticum (MG) genomic library constructed in plasmid pUC8. Based on the DNA sequence data of fMG-2, a pair of 25 base primers, designated amplification (Amp) left (L) and right (R) primers, was synthesized. When used in the polymerase chain reaction (PCR), the Amp L and R primers directed amplification of DNA of 16 MG strains yielding an expected 732-bp product, but did not amplify DNA of Escherichia coli, calf thymus, lambda phage, pUC8 plasmid, or 16 other species of avian mycoplasmas. As low as 10(-6) picogram of MG DNA, a fraction of the total chromosomal content of one cell, was detected following amplification by PCR. PCR amplification products were visualized by either ethidium bromide/ultraviolet exposure or hybridization with a 481-bp probe (fMG-3) prepared from the central region of fMG-2.
Nascimento
E R
ER
Department of Epidemiology and Preventive Medicine, School of Veterinary Medicine, University of California, Davis 95616.
Yamamoto
R
R
Herrick
K R
KR
Tait
R C
RC
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011200:Poultry; D011201:Poultry Diseases / Q000175:diagnosis*; D011237:Predictive Value of Tests
D004269:DNA, Bacterial