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Developmental Studies on the Laboratory diagnosis at an early stage of Mycoplasam pneumoniae-infection (mycoplasmal pneumonia)

Research Project

Project/Area Number 02557023
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 細菌学
Research InstitutionOkayama University

Principal Investigator

KANEMASA Yasuhiro  Okayama University Medical School, Department of Microbiology, Professor, 医学部, 教授 (80033059)

Co-Investigator(Kenkyū-buntansha) FUJII Mari  Okayama University Medical School Department of Microbiology, Research Associate, 医学部, 助手 (40212485)
MATSUSHITA Osamu  Okayama University Medical School Department of Microbiology, Research Associate, 医学部, 助手 (00209537)
KOTANI Nobuyuki  Okayama University Medical School, Department of Pediatrics, Assistant Professor, 医学部附属病院, 講師 (80231314)
KUNITOMI Taiji  Okayama University Medical School, Department of Pediatrics, Assistant Professor, 医学部附属病院, 講師 (10033292)
HIRAI Yoshikazu  Okayama University Medical School, Department of Microbiology, Associate Profess, 医学部, 助教授 (00127581)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1990: ¥4,000,000 (Direct Cost: ¥4,000,000)
KeywordsMycoplasma pneumonia / Mycoplasmal Pneumonia / Indirect Immunofluorescence Test / Latex Agglutination Test / DNA-probe Test / Respiratory Exudates / Laboratory Diagnosis / Throat Swabs / ラテックス凝集反応 / DNA-プロ-ブ / Mycoplasma pheumoniae / DNAプロ-ブ
Research Abstract

We developed an indirect immunofluorescence test (IF) and a latex agglutination test (LAT) for detection of Mycoplasma pneumonias in respiratory exudated as rapid diagnosis at an early stage of M. pneumoniae-infection. Further, IF and LAT were compared with DNA-probe test (DP) which was the only commercially available test for the rapid detection of the organism.
Firstly, we prepared polyclonal antibody specific to M. Pneumoniae. The antibody cross-react with many species of mycoplasmas, but not with normal human serum or with respiratory exudates from healthy person. Cross-reactivity of the antibody with species of mycoplasmas other than M. Renitalium was fully diminished when absorbed with horse serum and yeast extract. components of culture medium. -M. genitalium showed a cross-reaction in LAT with using the absorbed antibody and also in DP. However, the cross-reactivity with M. Renitalium, is unlikely to cause significant diagnostic confusion because the titer of M. genitalium was s … More ignificantly lower than., that of M. pneumonia, and M. genitalium is exclusively a genital tract colonizer. Therefore, IF and LAT with using the absorbed antibody, and DP are specific enough to be used for the detection of M. pneumonia in respiratory exudates.
The detection limit of IF was about 2 x 10^5 cfu/ml, that-, of LAT was 2 x 10^5 cfu/ml and that cif DP was 5 x 10^4 cfu/ml in vitro. DP had the highest sensitivity among three methods.
Among clinical specimens(throat smears)from patients with serologically confirmed M. pneumoniae-infection, 85.7% gave a positive test in IF. Among clinical specimens in which M. pneumoniae was detected by culture method, positive rate in IF was 73.3%, that in LAT was 63.3% and that in DP was 26.6%.
A effect of the incubation of M. pneumonia suspension was examined. The suspension and the mixture (M. Pneumoniae suspension and respiratory exudates) were incubated at 37 ゚C. An aliquot was removed with time course, and examinedin LAT and DP. The samples showed the same titer within three days in LAT. The titer (rate of sample-cpm to negativp- control-cpm) of each sample decreased remarkably within 5 h. It was considered that target SS'olecules in LAT and IF were accumulated in the pharyngeal portion. However, target molecule in DP (ribosomal RNA) was destructed much sooner, and the accumulation could not be expected. The reason f. or the low positive rate of DP in clinical'specimens may be due to the fast breakdown of target molecule.
Consequently, IF and LAT must be applicable to the detection of M. pneumoniae in respiratory exudates from the patients in clinical laboratories. Especially, LAT was recommended because the procedure of LAT can be completed within a half hour without complicated manner. though the positive rate of LAT in clinical specimens was slightly lower than that of IF. Now we are examinating a process of treating clinical samples for rising the detection limit of LAT. Less

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] 国富 泰二,小谷 信行,濃野 信,平井 義一,清野 佳紀: "蛍光抗体法によるマイコプラズマ肺炎の迅速診断" 小児科臨床. 43. 2288-2291 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 国富 泰二,小谷 信行,濃野 信,平井 義一,清野 佳紀: "間接蛍光抗体法による気管支喘息発作時のMycoplasma pneumoniae抗原の迅速検出" 小児科診療. 53. 2830-2833 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y.Hirai,J.Shiode, T.Masayoshi,Y.Kanemasa: "Application of Indirect Immunofluorescence Test for Detection of Mycoplasma pneumoniae in Respiratory Exudates" J.Clinical Microbiol.29(9). 2007-2012 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y.Hirai,T.Kanatani,M.Ono,O.Matsushita,Y.Kanemasa: "An Indirect Immunofluorescence Method for Detection of Mycoplasma hominis in Vaginal Smears" Microbiol.Immunol.35(10). 831-839 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T.Masayoshi,Y.Hirai,Y.Kanemasa: "A Latex Agglutination Test for the Detection of Mycoplasma pneumoniae in Respiratory Exudates: A Comparative Study with a Commercially Available DNA-probe Test" Microbiol.Immunol.36(2). (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Taiji kunitomi, Nobuyuki Kotani, Shin Nouno, Yoshikazu Hirai and Yoshiki Seino: "An indirect immunofluorescence test for rapid diagnosis of mycoplasmal pneumonia." Shounika Rinshou. 43(11). 2288-2291 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Taiji Kunitomi, Nobuyuki Kotani, Shin Nouno, Yoshikazu Hirai and Yoshiki Seino: "Rapid detection of Mycoplasma pneumonia in respiratory exudates by immunofluorescence at attack of bronchial asthma." Shounika Shinryou. 53(11). 2830-2833 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yoshikazu Hirai, Junji Shiode, Toyohisa Masayoshi and Yasuhiro Kanemasa: "Application of an indirect immunofluorescence test for detection of Mycoplasma pneumoniae in respiratory exudates." Journal of Clinical Microbiology. 29(9). 2007-2012 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yoshikazu Hirai, Tomohisa Kanatani, Masako Ono, Matsushita and Yasuhiro Kanemasa: "An indirect immunofluorescence method for detection of Mycoplasma hominis in vaginal smears." Microbiology and Immunology. 35(10). 831-839 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Toyohisa Masayoshi, Yoshikazu Hirai and Yasuhiro Kanemasa: "A latex agglutination test for the detection of Mycoplasma Pneumoniae in respiratory exudates : A comparative study with a commercially available DNA-probe test." Microbiology and Immunology. 36(2). (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y.Hirai,J.Shiode,T.Masayoshi,Y.Kanemasa: "Application of Indirect Immunofluorescence Test for Detection of Mycoplasma pneumoniae in Respiratory Exudates" J.Clinical Microbiol.29(9). 2007-2012 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Y.Hirai,T.Kanatani,M.Ono,O.Matsushita,Y.Kanemasa: "An Indirect Immunofluorescence Method for Detection of Mycoplasma hominis in Vaginal Smears" Microbiol.Immunol.35(10). 831-839 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] T.Masayoshi,Y.Hirai,Y.Kanemasa: "A Latex Agglutination Test for the Detection of Mycoplasma pneumoniae in Respiratory Exudates:A Comparative Study with a Commerciall Available DNA-Probe Test" Microbiol.Immunol.36(2). (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 国富 泰二,小谷 信行,濃野 信,平井 義一,清野 佳紀: "蛍光抗体法によるマイコプラズマ肺炎の迅速診断" 小児科臨床. 43. 2288-2291 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 国富 泰二,小谷 信行,濃野 信,平井 義一,清野 佳紀: "間接蛍光抗体法による気管支喘息発作時のMycoplasma Pneumoniae抗原の迅速検出" 小児科診療. 53. 2830-2833 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Y.Hirai,T.Masayoshi,J.Shiode,Y.Kanemasa: "Indirect Immunofluorescence Test for the Direct Detection of the Specific Antigen of Mycoplasma pneumoniae and its Application to Respiratory Exdates for Laboratory Diagnosis" J.clinicol microbiology.

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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