Polymerase chain reaction with a primer pair in the 16S-23S rRNA spacer region for detection of Mycoplasma pulmonis in clinical isolates.
To develop a diagnostic tool to identify Mycoplasma pulmonis (M. pulmonis) in clinical isolates, we developed a polymerase chain reaction (PCR) assay using primers specific for the 16S-23S rRNA intergenic spacer region (SR) of M. pulmonis. One pair of PCR primers reacted specifically with two reference strains of M. pulmonis tested and seven samples isolated from naturally infected rats. The primer pair did not produce PCR products of the correct size from any other rodent or human mycoplasmas or cellular DNA from rodent lungs. Specificity of the PCR assay was confirmed by Southern blotting with probe specific for the SR of M. pulmonis. The PCR assay for detection of M. pulmonis established in this study is suitable for diagnosis of M. pulmonis infection in clinical cases.
Takahashi-Omoe
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Department of Technical Support and Development, National Institute of Radiological Science, 4-9-1 Anagawa, Inage-ku, Chiba-shi 263-8555, Japan. h_omoe@nirs.go.jp
Omoe
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Matsushita
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Kobayashi
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Yamamoto
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D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern / Q000662:veterinary; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics*; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D045803:Mycoplasma pulmonis / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012313:RNA / Q000737:chemistry / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D012376:Rodent Diseases / Q000382:microbiology*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; C034477:RNA primers; D012336:RNA, Ribosomal, 16S; D012313:RNA