Skip to main content
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1996 Oct;49(10):861–863. doi: 10.1136/jcp.49.10.861

Sensitive and universal method for microbial DNA extraction from blood products.

N Golbang 1, J P Burnie 1, P E Klapper 1, A Bostock 1, P Williamson 1
PMCID: PMC500787  PMID: 8943759

Abstract

A new method of extracting bacterial and yeast DNA from blood products dependent on guanidinium thiocyanate acid extraction and proteinase K treatment is described. In spiked samples the sensitivity per 0.1 ml of serum and blood, respectively, was 26 and 150 colony forming units (cfu) for Escherichia coli, 80 and 120 cfu for Staphylococcus aureus and 20 and 26 cfu for Candida albicans. This compared well with existing methodologies, worked on limited clinical samples and was not pathogen specific.

Full text

PDF
861

Images in this article

Selected References

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

  1. Attal J., Cajero-Juarez M., Houdebine L. M. A simple method of DNA extraction from whole tissues and blood using glass powder for detection of transgenic animals by PCR. Transgenic Res. 1995 Mar;4(2):149–150. doi: 10.1007/BF01969417. [DOI] [PubMed] [Google Scholar]
  2. Burgener-Kairuz P., Zuber J. P., Jaunin P., Buchman T. G., Bille J., Rossier M. Rapid detection and identification of Candida albicans and Torulopsis (Candida) glabrata in clinical specimens by species-specific nested PCR amplification of a cytochrome P-450 lanosterol-alpha-demethylase (L1A1) gene fragment. J Clin Microbiol. 1994 Aug;32(8):1902–1907. doi: 10.1128/jcm.32.8.1902-1907.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Casas I., Powell L., Klapper P. E., Cleator G. M. New method for the extraction of viral RNA and DNA from cerebrospinal fluid for use in the polymerase chain reaction assay. J Virol Methods. 1995 May;53(1):25–36. doi: 10.1016/0166-0934(94)00173-e. [DOI] [PubMed] [Google Scholar]
  4. Chryssanthou E., Andersson B., Petrini B., Löfdahl S., Tollemar J. Detection of Candida albicans DNA in serum by polymerase chain reaction. Scand J Infect Dis. 1994;26(4):479–485. doi: 10.3109/00365549409008623. [DOI] [PubMed] [Google Scholar]
  5. Condos R., McClune A., Rom W. N., Schluger N. W. Peripheral-blood-based PCR assay to identify patients with active pulmonary tuberculosis. Lancet. 1996 Apr 20;347(9008):1082–1085. doi: 10.1016/s0140-6736(96)90281-0. [DOI] [PubMed] [Google Scholar]
  6. Crampin A. C., Matthews R. C. Application of the polymerase chain reaction to the diagnosis of candidosis by amplification of an HSP 90 gene fragment. J Med Microbiol. 1993 Sep;39(3):233–238. doi: 10.1099/00222615-39-3-233. [DOI] [PubMed] [Google Scholar]
  7. Espy M. J., Patel R., Paya C. V., Smith T. F. Comparison of three methods for extraction of viral nucleic acids from blood cultures. J Clin Microbiol. 1995 Jan;33(1):41–44. doi: 10.1128/jcm.33.1.41-44.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fujita S., Lasker B. A., Lott T. J., Reiss E., Morrison C. J. Microtitration plate enzyme immunoassay to detect PCR-amplified DNA from Candida species in blood. J Clin Microbiol. 1995 Apr;33(4):962–967. doi: 10.1128/jcm.33.4.962-967.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greisen K., Loeffelholz M., Purohit A., Leong D. PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid. J Clin Microbiol. 1994 Feb;32(2):335–351. doi: 10.1128/jcm.32.2.335-351.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jones J. M. Laboratory diagnosis of invasive candidiasis. Clin Microbiol Rev. 1990 Jan;3(1):32–45. doi: 10.1128/cmr.3.1.32. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kan V. L. Polymerase chain reaction for the diagnosis of candidemia. J Infect Dis. 1993 Sep;168(3):779–783. doi: 10.1093/infdis/168.3.779. [DOI] [PubMed] [Google Scholar]
  12. Kanai N., Fujii T., Saito K., Tokoyama T. Rapid and simple method for preparation of genomic DNA from easily obtainable clotted blood. J Clin Pathol. 1994 Nov;47(11):1043–1044. doi: 10.1136/jcp.47.11.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Miyakawa Y., Mabuchi T., Fukazawa Y. New method for detection of Candida albicans in human blood by polymerase chain reaction. J Clin Microbiol. 1993 Dec;31(12):3344–3347. doi: 10.1128/jcm.31.12.3344-3347.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rudolph K. M., Parkinson A. J., Black C. M., Mayer L. W. Evaluation of polymerase chain reaction for diagnosis of pneumococcal pneumonia. J Clin Microbiol. 1993 Oct;31(10):2661–2666. doi: 10.1128/jcm.31.10.2661-2666.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rådström P., Bäckman A., Qian N., Kragsbjerg P., Påhlson C., Olcén P. Detection of bacterial DNA in cerebrospinal fluid by an assay for simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and streptococci using a seminested PCR strategy. J Clin Microbiol. 1994 Nov;32(11):2738–2744. doi: 10.1128/jcm.32.11.2738-2744.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schaberg D. R., Culver D. H., Gaynes R. P. Major trends in the microbial etiology of nosocomial infection. Am J Med. 1991 Sep 16;91(3B):72S–75S. doi: 10.1016/0002-9343(91)90346-y. [DOI] [PubMed] [Google Scholar]
  17. Siafakas N., Burnett L., Bennetts B., Proos A. Nonenzymatic extraction of DNA from blood collected into serum separator tubes. Clin Chem. 1995 Jul;41(7):1045–1046. [PubMed] [Google Scholar]
  18. Song J. H., Cho H., Park M. Y., Na D. S., Moon H. B., Pai C. H. Detection of Salmonella typhi in the blood of patients with typhoid fever by polymerase chain reaction. J Clin Microbiol. 1993 Jun;31(6):1439–1443. doi: 10.1128/jcm.31.6.1439-1443.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ubukata K., Nakagami S., Nitta A., Yamane A., Kawakami S., Sugiura M., Konno M. Rapid detection of the mecA gene in methicillin-resistant staphylococci by enzymatic detection of polymerase chain reaction products. J Clin Microbiol. 1992 Jul;30(7):1728–1733. doi: 10.1128/jcm.30.7.1728-1733.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zhang Y., Isaacman D. J., Wadowsky R. M., Rydquist-White J., Post J. C., Ehrlich G. D. Detection of Streptococcus pneumoniae in whole blood by PCR. J Clin Microbiol. 1995 Mar;33(3):596–601. doi: 10.1128/jcm.33.3.596-601.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. van Deventer A. J., Goessens W. H., van Belkum A., van Vliet H. J., van Etten E. W., Verbrugh H. A. Improved detection of Candida albicans by PCR in blood of neutropenic mice with systemic candidiasis. J Clin Microbiol. 1995 Mar;33(3):625–628. doi: 10.1128/jcm.33.3.625-628.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES