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Peptoniphilus mikwangii -specific quantitative real-time polymerase chain reaction primers

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초록

The purpose of this study was to develop Peptoniphilus mikwangii -specific quantitative real-time polymerase chain reaction (qPCR) primers based on the 16S ribosomal RNA (16S rDNA) gene. The specificity of the primers was determined by conventional PCR using 29 strains of 27 oral bacterial species including P. mikwangii . The sensitivity of the primers was determined by qPCR using the purified genomic DNA of P. mikwangii KCOM 1628 T (40 ng to 4 fg). The data showed that the qPCR primers (RTB134-F4/RTB134-R4) could detect P. mikwangii strains exclusively and as little as 40 fg of the genomic DNA of P. mikwangii KCOM 1628 T . These results suggest that the developed qPCR primer pair can be useful for detecting P. mikwangii in epidemiological studies of oral bacterial infectious diseases.

Abstract

The purpose of this study was to develop Peptoniphilus mikwangii -specific quantitative real-time polymerase chain reaction (qPCR) primers based on the 16S ribosomal RNA (16S rDNA) gene. The specificity of the primers was determined by conventional PCR using 29 strains of 27 oral bacterial species including P. mikwangii . The sensitivity of the primers was determined by qPCR using the purified genomic DNA of P. mikwangii KCOM 1628 T (40 ng to 4 fg). The data showed that the qPCR primers (RTB134-F4/RTB134-R4) could detect P. mikwangii strains exclusively and as little as 40 fg of the genomic DNA of P. mikwangii KCOM 1628 T . These results suggest that the developed qPCR primer pair can be useful for detecting P. mikwangii in epidemiological studies of oral bacterial infectious diseases.

참고문헌 (9)

  1. Eugene Cho, "Peptoniphilus mikwangii sp. nov., Isolated from a Clinical Specimen of Human Origin", Current Microbiology, 70(2), pp.260~266 (2015), Springer Science and Business Media LLC[DOI]
  2. 박순낭, "Development of Species-specific PCR Primers for Detecting Peptoniphilus mikwangii", International Journal of Oral Biology, 42(3), pp.143~147 (2017), 대한구강생물학회
  3. Krieg NR, "Bergey’s manual of systematic bacteriology", pp.33~38 (2001), Springer
  4. Woese CR, "Bacterial evolution", Microbiol Rev, 51, pp.221~271 (1987)[DOI]
  5. Soon-Nang Park, "Development of quantitative real-time PCR primers for detecting 42 oral bacterial species", Archives of Microbiology, 195(7), pp.473~482 (2013), Springer Science and Business Media LLC[DOI]
  6. Wilson K, "Current protocols in molecular biology", pp.241~245 (1990), Greene Publ. Assoc. and Wiley Interscience
  7. 박순낭, "Development of Quantitative Real-Time PCR Primers for Detection of Prevotella intermedia", International Journal of Oral Biology, 40(4), pp.205~210 (2015), 대한구강생물학회[DOI]
  8. E. Cho, "Draft Genome Sequence of the Novel Peptoniphilus sp. Strain ChDC B134, Isolated from a Human Periapical Abscess Lesion", Genome Announcements, 1(5), pp.e00822-13~ (2013), American Society for Microbiology[DOI]
  9. 박순낭, "Development of Porphyromonas gingivalis-Specific Quantitative Real-Time PCR Primers Based on the Nucleotide Sequence of rpoB", The Journal of Microbiology, 49(2), pp.315~319 (2011), 한국미생물학회

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