Hostname: page-component-78c5997874-v9fdk Total loading time: 0 Render date: 2024-11-14T16:46:43.767Z Has data issue: false hasContentIssue false
  • English
  • Français

Laboratory diagnosis of Mycoplasma pneumoniae infection. 4. Antigen capture and PCR-gene amplification for detection of the mycoplasma: problems of clinical correlation

Published online by Cambridge University Press:  15 May 2009

J. Williamson ,
B. P. Marmion ,
D. A. Worswick ,
T. -W. Kok ,
G. Tannock ,
R. Herd  and
R. J. Harris
J. Williamson
Affiliation:
School of Pharmacy and Medical Laboratory Science, University of South Australia, Adelaide, South Australia
B. P. Marmion*
Affiliation:
Department of Pathology, University of Adelaide, South Australia Division of Medical Virology, Institute of Medical and Veterinary Science, Adelaide, South Australia
D. A. Worswick
Affiliation:
Division of Medical Virology, Institute of Medical and Veterinary Science, Adelaide, South Australia
T. -W. Kok
Affiliation:
Division of Medical Virology, Institute of Medical and Veterinary Science, Adelaide, South Australia
G. Tannock
Affiliation:
Faculty of Medicine, The University of Newcastle, Newcastle, New South Wales
R. Herd
Affiliation:
Faculty of Medicine, The University of Newcastle, Newcastle, New South Wales
R. J. Harris
Affiliation:
School of Pharmacy and Medical Laboratory Science, University of South Australia, Adelaide, South Australia
*
*Please address correspondence and reprint requests to: Professor B. P. Marmion, Department of Pathology, University of Adelaide, North Terrace, Adelaide, South Australia 5000.
Rights & Permissions [Opens in a new window]

Summary

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Direct detection assays for Mycoplasma pneumoniae were established by PCR amplification of short sequences within the foot protein/adhesin (P1) gene and the 16S ribosomal RNA gene.

Specificity and sensitivity was excellent, no hybridization was observed with M. genitalium and other human Mycoplasma species. In nose and throat washings from subjects with respiratory infection a pattern of high counts (c.f.u./ml) of M. pneumoniae (deduced from the amount of amplified PCR product), and a positive antigen capture assay, was found in 83% of subjects with serological evidence of current infection with M. pneumoniae.

A small proportion of subjects with serological patterns suggesting infection in the more distant past had positive PCR assays. This was considered to represent either persistence of the organism from a previous infection or perhaps transient carriage during a reinfection, without substantial change in antibody response.

PCR-based assay of M. pneumoniae offers a powerful, rapid, and sensitive substitute for culture of the mycoplasma. Antigen capture, while less sensitive than PCR, offers the advantage that it is more often positive with samples from current infection and requires less stringent laboratory organization to contain false positive results. We conclude however that the laboratory diagnosis of a chosen clinical episode should not rest on the PCR or Ag-EIA assays alone, but must also include antibody assays to confirm whether infection is current or represents persistence from past exposure.

Type
Research Article
Information
Epidemiology & Infection , Volume 109 , Issue 3 , December 1992 , pp. 519 - 537
Copyright
Copyright © Cambridge University Press 1992

References

REFERENCES

1.Kok, T-W, Varkanis, G, Marmion, BP, Martin, J, Esterman, A. Laboratory diagnosis of Mycoplasma pneumoniae infection. 1. Direct detection of antigen in respiratory exudates by enzyme immunoassay. Epidemiol Infect 1988; 101: 669–84.CrossRefGoogle ScholarPubMed
2.Harris, R, Marmion, BP, Varkanis, G, Kok, T, Lunn, B, Martin, J. Laboratory diagnosis of Mycoplasma pneumoniae infection. 2. Comparison of methods for the direct detection of specific antigen or nucleic acid sequences in respiratory exudates. Epidemiol Infect 1988; 101: 685–94.CrossRefGoogle ScholarPubMed
3.Saiki, RK, Gelfand, DH, Stoffel, S, et al. . Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988; 239: 487–91.CrossRefGoogle ScholarPubMed
4.Bernet, C, Garret, M, de Barbyrac, B, Bebear, C, Bonnet, J. Detection of M. pneumoniae by using the polymerase chain reaction. J Clin Microbiol 1989; 27: 2492–6.CrossRefGoogle ScholarPubMed
5.Jensen, SJ, Sondergard, J, Uldum, SA, Lind, K. Detection of M. pneumoniae in simulated clinical samples by polymerase chain reaction. APMIS 1989; 97: 1046–8.CrossRefGoogle ScholarPubMed
6.Jensen, JS, Uldum, SA, Sondergard-Andersen, J, Vuust, J, Lind, K. Polymerase chain reaction for detection of Mycoplasma genitalium in clinical samples. J Clin Micro 1991; 29: 46–50.CrossRefGoogle ScholarPubMed
7.Taylor-Robinson, D. Genital mycoplasma infections. Clinics Lab Med 1989; 9: 501–23.CrossRefGoogle ScholarPubMed
8.Taylor-Robinson, D, Smith, GR. Mycoplasma diseases of man and animals In: Smith, GR, Easmon, CSF, eds. Topley and Wilson's principles of bacteriology, vol. 3. Bacterial diseases, 8th edn. London: Edward Arnold, 1990; 657–72.Google Scholar
9.Palmer, HM, Gilroy, CB, Furr, PM, Taylor-Robinson, D. Development and evaluation of the polymerase chain reaction to detect Mycoplasma genitalium. FEMS Microbiol Letts 1991; 77: 199–204.CrossRefGoogle Scholar
10.Palmer, HM, Gilroy, CB, Claydon, EJ, Taylor-Robinson, D. Detection of Mycoplasma genitalium in the genitourinary tract of women by the polymerase chain reaction. Int J STD AIDS 1991; 2: 261–3.CrossRefGoogle ScholarPubMed
11.Tully, JG, Whitcomb, RF, Clark, HF, Williamson, DL. Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new spiroplasma. Science 1977; 195: 892–4.CrossRefGoogle ScholarPubMed
12.Marmion, BP. Mycoplasma: acholeplasma: ureaplasma. In: Collee, JG, Duguid, JP, Fraser, AG, Marmion, BP, eds. Mackie & McCartney' s practical medical microbiology, 13th edn. Edinburgh: Churchill Livingstone, 1989; 745–68.Google Scholar
13.Kok, TW, Mamion, BP, Varkanis, G, Worswick, DA, Martin, J. Laboratory diagnosis of Mycoplasma pneumoniae infection. 3. Detection of IgM antibodies to M. pneumoniae by a modified indirect haemagglutination test. Epidemiol Infect 1989; 103: 613–23.CrossRefGoogle Scholar
14.Sriprakash, KS, Hartas, J. Hairpin extension. A general method for the improvement of sensitivity of oligonucleotide probes. Gene Anal Techn 1989; 6: 29–32.CrossRefGoogle ScholarPubMed
15.Inamine, JM, Denny, TP, Loechel, S, et al. . Nucleotide sequence of the P1 attachment-protein of Mycoplasma pneumoniae. Gene 1988; 64: 217–29.CrossRefGoogle ScholarPubMed
16.Dallo, SF, Chavoya, A, Su, C-J, Baseman, JB. DNA and protein sequence homologies between the adhesins of Mycoplasma genitalium and Mycoplasma pneumoniae. Infect Immun. 1989; 57: 1059–65.CrossRefGoogle ScholarPubMed
17.Weisbergh, WG, Tully, JG, Rose, DL, et al. . A phylogenetic analysis of the mycoplasmas: Basis for their classification. J Bacteriol 1989; 171: 6455–67.CrossRefGoogle Scholar
18.Su, C-J, Tryon, VV, Baseman, JB. Cloning and sequence analysis of cytadhesin P1 gene from Mycoplasma pneumoniae. Infect Immun 1987; 55: 3023–9.CrossRefGoogle ScholarPubMed
19.Hu, PC, Powell, DA, Albright, F, Gardner, DE, Collier, AM, Clyde, WA. A solid-phase radioimmunoassay for detection of antibodies against Mycoplasma pneumoniae. J Clin Lab Immunol 1983; 11: 209–13.Google ScholarPubMed
20.Lind, K, Lindhart, BO, Schutten, HJ, Blom, J, Christiansen, D. Serological cross-reactions between Mycoplasma genitalium and Mycoplasma pneumoniae. J Clin Microbiol 1984; 20: 1036–43.CrossRefGoogle ScholarPubMed
21.Yogev, D, Razin, S, Common deoxyribonucleic acid sequences in Mycoplasma genitalium and Mycoplasma pneumoniae genomes. Int J Sys Bacteriol 1986; 36: 426–30.CrossRefGoogle Scholar
22.Baseman, JB, Dallo, SF, Tully, JG, Rose, DL. Isolation and characterisation of M. genitalium strains from the human respiratory tract. J Clin Microbiol 1988; 26: 2266–9.CrossRefGoogle ScholarPubMed
23.Couch, RB, Cate, TR, Chanock, RM. Infection with artificially propagated Eaton Agent (Mycoplasma pneumoniae). JAMA 1964; 187: 146–51.CrossRefGoogle ScholarPubMed