Submitted: 23 Oct 2013
Revised: 10 Nov 2013
Accepted: 25 Nov 2013
First published online: 05 Oct 2016
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Int J Enteric Pathog. 2014;2(1): e15595.
doi: 10.17795/ijep15595
  Abstract View: 1001
  PDF Download: 569

Research Article

Comparing the Accuracy Rate of Two Different Universal Primers in Enteric Pathogen Diagnosis From Blood

Esmaeil Soleimani 1, Hamidreza Honarmand 2 * , Iraj Nikokar 3, Zinab Falakian 3

1 Department of Microbiology, Islamic Azad University, Lahijan branch, Lahijan, I.R. Iran
2 Department of Microbiology, school of Medicine, Guilan University of Medical Sciences, Rasht, I.R. Iran
3 School of Nursery, Midwifery and Paramedicine, Guilan University of Medical Sciences, Langroud, I.R. Iran
*Corresponding author: Hamidreza Honarmand, Department of Microbiology, school of Medicine, Guilan University of Medical Sciences, Rasht, I.R. Iran. Tel: +98-9111434698, E-mail: honarmand@gums.ac. Email: irhonarmand.3@gmail.com

Abstract

Background: Detecting enteric bacteria in blood by culture is a slow assay with low accuracy rate. PCR might be a suitable alternative assay but as several species can cause bacteremia, it is necessary to use universal primers.

Objectives: In this study we evaluated and compared two pairs of universal primers in detecting four enteric bacteria in blood, which are common causes of bacteremia in human.

Materials and Methods: Standard strains of E. faecalis, S. typhi, E. coli, and S. Aeruginosa, were used in this study. A serially diluted bacterial suspension of all strains was made for inoculation to four sets of defibrinated sheep blood which were used to prepare blood specimens with different bacterial contents for performing routine assay and PCR. PCR was performed using two different universal primers designed from two ribosomal genes, 16sr RNA and 23sr RNA.

Results: PCR with 16sr RNA universal primer showed more accuracy rate than both blood culture and PCR with 23sr RNA universal primer. Mean time for performing PCR assay and blood culture was eight and 48 hours, respectively.

Conclusions: Both PCR with 16sr RNA and 23sr RNA universal primers have more accuracy rate than blood culture and are faster in detection of bacteremia. PCR with 16sr RNA universal primer is more accurate than both PCR with 16sr RNA universal primer and blood culture for diagnosis of bacteremia.

Implication for health policy/practice/research/medical education:

Detecting bacteria in blood by culture is a slow assay with low accuracy rate. PCR might be a suitable alternative assay; however, several species can cause bacteremia and it is necessary to use the universal primers. In this study, two pairs of universal primers for detecting seven bacteria in blood were evaluated and compared, which are common causes of human bacteremia. We found that both PCR with 16sr RNA and 23sr RNA universal primers have high accuracy rate and are faster than blood culture in detection of bacteremia. PCR with16Sr RNA universal primer is more accurate in diagnosis of bacteremia than both PCR with 16sr RNA universal primer and culture.

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