Activity and DNA contamination of commercial polymerase chain reaction reagents for the universal 16S rDNA real-time polymerase chain reaction detection of bacterial pathogens in blood

作者：Muehl Helge; Kochem Anna Julia; Disque Claudia; Sakka Samir G^*

来源：Diagnostic Microbiology and Infectious Disease, 2010, 66(1): 41-49.

DOI：10.1016/j.diagmicrobio.2008.07.011

• Tag polymerase activity
• DNA-free PCR reagent
• Universal 16S rDNA PCR
• Bacteremia
• Sensitivity of detection in blood

Universal 16S rRNA gene polymerase chain reaction (PCR) is a promising means of detecting bacteremia. Among other factors, the PCR reagents play a prominent role for obtaining a high sensitivity of detection. The reagents are ideally optimized with respect to the amplifying activity and absence of contaminating DNA. In this study, it was shown in a universal 16S rDNA real-time PCR assay that commercial PCR reagents can vary greatly among each other in these characters. Only 1 of the 5 reagents tested met the criteria of sensitive detection of pathogen DNA with a minimum of false-positive results. The reagent was validated by the detection of pathogens at low titers using bacterial DNA extracted from blood that was spiked with various Gram-positive and Gram-negative bacteria.

• 出版日期2010-1