Abstract
Chronic polymicrobial infections represent diagnostic challenges for both molecular and culture methods. Chronic wounds, inducing chronic pain and reducing the mobility of individuals, have a notable effect on the quality of life. At any given time, 1% of the population is usually affected and, therefore, multiple treatments are also required. Identification of the etiology of such infections facilitates the appropriate treatment. In this study, 20 wound samples were collected from 20 patients with suspected wound infection. The panel of the current assay targets 12 clinically relevant aerobic pathogens, commonly associated with chronic wound infection. Microbial wound infections were detected by both usual polymerase chain reaction (PCR) and subsequently testing using the DNA chip. In the current study, the results of culture-free bacterial identification using the two methods of DNA analyses were compared. By molecular detection using PCR, seven different bacterial species were identified: Citrobacter spp. (100%), Enterobacter spp. (100%), Klebsiella pneumoniae (100%), Pseudomonas aeruginosa (90%), Proteus spp. (80%), Escherichia coli (60%), and Staphylococcus aureus (10%). Mixed microbial infections were detected in all samples indicating four, five, or six different bacteria, identified in each sample. Microarray detection in comparison to PCR indicated 100% matching. These results demonstrate the possibility of fast identification of wound infection pathogens even in a mixed culture in a very short time, which in turn facilitates the proceeding steps for proper treatment.
Highlights
Microarray detection in comparison to polymerase chain reaction (PCR) indicated 100% matching. These results demonstrate the possibility of fast identification of wound infection pathogens even in a mixed culture in a very short time, which in turn facilitates the proceeding steps for proper treatment
One of the most common health problems is usually associated with wound infection (Giacometti et al, 2000), commonly induced by the entry of the bacteria through contamination of breached skin
The pressure induced on both patients and the medical system, efforts have been made with the aim of managing wound infections and their associated pathological conditions, which in turn could improve the quality of life and increase life expectancy
Summary
One of the most common health problems is usually associated with wound infection (Giacometti et al, 2000), commonly induced by the entry of the bacteria through contamination of breached skin. Universal primers for 16S rRNA genes are used, followed by specific identification approaches such as polymerase chain reaction (PCR) (Hill et al, 2003) and denaturing gradient gel electrophoresis and sequencing (Dowd et al, 2008a, 2008b; Hill et al, 2003; Price et al, 2009) These methods exhibited successfulness in the detection of bacteria in burn wounds (Pirnay et al, 2000), blood (Rothman et al, 2002; Wellinghausen et al, 2004; Yang et al, 2002), cerebrospinal fluid (Poppert et al, 2005), joint fluid (Yang et al, 2008), and heart tissue (Breitkopf et al, 2005). For developing new therapeutic strategies, the use of these techniques speeds up the early detection of infecting microorganisms assisting clinicians and pathologists to find out the appropriate treatment of hospitalized patients
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