Skip to main content
PMC home page
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1992 Oct;45(10):910–913. doi: 10.1136/jcp.45.10.910

Use of the polymerase chain reaction to detect Toxoplasma gondii in human blood samples.

D O Ho-Yen 1, A W Joss 1, A H Balfour 1, E T Smyth 1, D Baird 1, J M Chatterton 1
PMCID: PMC495065  PMID: 1430262

Abstract

AIMS: To assess the value of detecting Toxoplasma gondii in human blood samples using the polymerase chain reaction (PCR). METHODS: Blood samples in lithium heparin were examined from 34 patients with suspected toxoplasmosis, and six healthy volunteers with or without the addition of doubling dilutions of toxoplasma tachyzoites. Products of a nested PCR, using oligonucleotide primers of the B1 gene, were analysed by electrophoresis and stained by ethidium bromide. The primary product was 194 base pairs in length; the nested products were 160 or 97 base pairs. RESULTS: When toxoplasma tachyzoites were added to the leucocytes of six different volunteers, eight to 16 parasites were detected by nested PCR in one experiment and one to four parasites in eight experiments. All nine experiments were negative in samples without added tachyzoites. Of 34 patients, PCR was negative in 13 with recent lymphadenopathy; nine with persisting IgM, including two pregnant patients; four with reactivated infection due to immunodeficiency; and five with no evidence of active infection. Positive PCR results were found in three patients with reactivated infection. There was only one discrepancy between PCR and animal culture results; this was in an immunocompromised patient with a positive PCR and negative culture. CONCLUSIONS: The PCR technique was rapid, sensitive, and specific in human blood samples. Negative PCR results in patients with persisting IgM suggested that the fetus was not at risk, or that treatment was not indicated in myalgic encephalomyelitis-like illness. PCR results in immunocompromised patients permitted appropriate management--no treatment if negative, treatment if positive.

Full text

PDF
910

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burg J. L., Grover C. M., Pouletty P., Boothroyd J. C. Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction. J Clin Microbiol. 1989 Aug;27(8):1787–1792. doi: 10.1128/jcm.27.8.1787-1792.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Hill A. B. A voice from the past. Public Health. 1988 Sep;102(5):407–407. doi: 10.1016/s0033-3506(88)80077-5. [DOI] [PubMed] [Google Scholar]
  3. Ho-Yen D. O. Toxoplasmosis in humans: discussion paper. J R Soc Med. 1990 Sep;83(9):571–572. doi: 10.1177/014107689008300913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Holliman R. E. Toxoplasmosis and the acquired immune deficiency syndrome. J Infect. 1988 Mar;16(2):121–128. doi: 10.1016/s0163-4453(88)93847-9. [DOI] [PubMed] [Google Scholar]
  5. Joss A. W., Skinner L. J., Chatterton J. M., Cubie H. A., Pryde J. F., Campbell J. D. Toxoplasmosis: effectiveness of enzyme immunoassay screening. Med Lab Sci. 1989 Apr;46(2):107–112. [PubMed] [Google Scholar]
  6. Joss A. W., Skinner L. J., Moir I. L., Chatterton J. M., Williams H., Ho-Yen D. O. Biotin-labelled antigen screening test for toxoplasma IgM antibody. J Clin Pathol. 1989 Feb;42(2):206–209. doi: 10.1136/jcp.42.2.206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Payne R. A., Joynson D. H., Balfour A. H., Harford J. P., Fleck D. G., Mythen M., Saunders R. J. Public Health Laboratory Service enzyme linked immunosorbent assay for detecting Toxoplasma specific IgM antibody. J Clin Pathol. 1987 Mar;40(3):276–281. doi: 10.1136/jcp.40.3.276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ruskin J., Remington J. S. Toxoplasmosis in the compromised host. Ann Intern Med. 1976 Feb;84(2):193–199. doi: 10.7326/0003-4819-84-2-193. [DOI] [PubMed] [Google Scholar]
  9. Savva D., Morris J. C., Johnson J. D., Holliman R. E. Polymerase chain reaction for detection of Toxoplasma gondii. J Med Microbiol. 1990 May;32(1):25–31. doi: 10.1099/00222615-32-1-25. [DOI] [PubMed] [Google Scholar]
  10. Skinner L. J., Chatterton J. M., Joss A. W., Moir I. L., Ho-Yen D. O. The use of an IgM immunosorbent agglutination assay to diagnose congenital toxoplasmosis. J Med Microbiol. 1989 Feb;28(2):125–128. doi: 10.1099/00222615-28-2-125. [DOI] [PubMed] [Google Scholar]
  11. Suzuki Y., Israelski D. M., Dannemann B. R., Stepick-Biek P., Thulliez P., Remington J. S. Diagnosis of toxoplasmic encephalitis in patients with acquired immunodeficiency syndrome by using a new serologic method. J Clin Microbiol. 1988 Dec;26(12):2541–2543. doi: 10.1128/jcm.26.12.2541-2543.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Weiss L. M., Udem S. A., Salgo M., Tanowitz H. B., Wittner M. Sensitive and specific detection of toxoplasma DNA in an experimental murine model: use of Toxoplasma gondii-specific cDNA and the polymerase chain reaction. J Infect Dis. 1991 Jan;163(1):180–186. doi: 10.1093/infdis/163.1.180. [DOI] [PubMed] [Google Scholar]
  13. van de Ven E., Melchers W., Galama J., Camps W., Meuwissen J. Identification of Toxoplasma gondii infections by BI gene amplification. J Clin Microbiol. 1991 Oct;29(10):2120–2124. doi: 10.1128/jcm.29.10.2120-2124.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES