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2013 | 60 | 4 | 603-606
Article title

Evaluation of the activity of thermostable DNA polymerases in the presence of heme, as a key inhibitor in the real time PCR method in diagnostics of sepsis

Content
Title variants
Languages of publication
EN
Abstracts
EN
The study aim was evaluation of the usefulness of several thermostable DNA polymerases in real time PCR conducted in the presence of the heme. Our study had the advantage of testing several different polymerases, one of which proved to be the least sensitive to heme activity. We also found that there is no need of supplementing the reaction mixture with protective substances like BSA. Selection of the appropriate polymerase can increase the efficiency of the PCR reaction which is very important for diagnosis of sepsis and for other analyses performed on DNA template isolated from the blood.
Keywords
Publisher

Year
Volume
60
Issue
4
Pages
603-606
Physical description
Dates
published
2013
received
2013-06-06
revised
2013-10-08
accepted
2013-11-12
(unknown)
2013-12-17
Contributors
  • Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
  • Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
  • Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
  • Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
  • Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
References
  • Abu Al-Soud W, Radstrom P (1998) Capacity of nine thermostable DNA Polymerases to Mediate DNA Amplification in the Presence of PCR - Inhibiting Samples. Appl Environ Microbiol 64: 3748-3753.
  • Abu Al-Soud W, Radstrom P (2001) Purification and Characterization of PCR-Inhibitory Components in Blood Cells. J Clin Microbiol 39: 485-493.
  • Abu Al-Soud W, Lantz P, Backman A, Olcen P, Radstorm P (1998) A sample preparation method which facilitates detection of bacteria in blood cultures by the polymerase chain reaction. J Microbiol Methods 32: 217-224.
  • Akane A, Matsubara K, Nakamura H, Takahashi S, Kimura K (1994) Identification of the Heme Compound Copurified with Deoxyribonucleic Acid (DNA) from Bloodstains a Major Inhibitor of Polymerase Chain Reaction Amplification. J Forensic Sci 39: 362-372.
  • Chiba N, Murayama SY, Morozumi M, Nakayama E, Okada T, Iwata S, Sunakawa K, Ubukata K (2009) Rapid detection of eight causative pathogens for the diagnosis of bacterial meningitis by real-time PCR. J Infect Chemother 15: 92-98.
  • Klouche M, Schroder U (2008) Rapid Methods for Diagnosis of Bloodstream Infections. Clin Chem Lab Med 46: 888-908.
  • Kreader CA (1996) Relief of Amplification Inhibition in PCR with Bovine Serum Albumin or T4 Gene 32 Protein. Appl Environ Microbiol 62: 1102-1106.
  • Lombardo ME, Araujo LS, Ciccarelli AB, Batlle A (2005) A Spectrophotometric Method for Estimating Hemin in Biological Systems. Anal Biochem 341: 199-203.
  • Opel KL, Chung D, McCord BR (2010) A Study of PCR Inhibition Mechanisms Using Real Time PCR. J Forensic Sci 55: 25-33.
  • Valle Jr DL, Andrade JI, Cabrera EC, Rivera WL (2010) Evaluation of Buffy Coat 16s rRNA PCR, Buffy Coat Culture and Whole Blood PCR for Detection of Bacteraemia. Mem Inst Oswaldo Cruz 105: 117–122.
Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv60p603kz
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