PCR Technique for the Microbial Analysis of Inanimate Hospital Environment

Abstract

Discipline of molecular ecology and molecular techniques such as polymerase chain reaction PCR ofers a possibility to study and reveal the microbial diversity in environmental setings with complicated mixed communities, non-culturable organisms, interfering contaminants and low levels of target DNA. Hospital environment represents a new ecological niche for clinically important nosocomial pathogens and antibiotic-resistant microorganisms, which have been commonly found on various hospital surfaces. Accurate characterization of microbial communities depends on several factors, starting with sample collection and conditional enrichment step. In the step of nucleic acid isolation and puriication, the DNA, as a dominant signature molecule, is extracted followed by removing co-extracted impurities. PCR target sequences are often 16S rDNA gene, functional gene probes or species-speciic probes, depending on the objective of the study. Furthermore, properly prepared PCR amplicons can serve as a basis for characterizing microbial community. The PCR technique is a powerful tool for the analysis of microbial diversity of environmental ecosystems. In a hospital environment, advantages of detecting pathogens and antibiotic-resistant bacteria need to be pointed out.Keywords: microorganisms, hospital environment, inanimate surfaces, DNA extraction, PCR . IntroductionCharacteristics of the hospital environments are very speciic where inanimate environment can be colonized with a wide range of microorganisms [1,2]. The cultured microorganisms represent only a small fraction of natural microbial communities, hence the microbial diversity in terms of species richness and species abundance is grossly underestimated [3]. Therefore, the discipline of molecular ecology and molecular techniques such as polymerase chain reaction PCR ofers a possibility to study and reveal the real-microbial population complexity and a possibility to overcome limitations of culture-based approaches. Due to the power of the PCR to amplify small amounts of DNA, organisms occurring in small numbers in an environment are now detectable [3]. Special challenges in environmental setings are complicated mixed communities, interfering contaminants and low levels of target DNA [4]. The speciics of environmental samples are low to medium concentration of target cells, low sample homogeneity and high degree of PCR inhibition [5].Many types of pathogenic microorganisms have been found on various common hospital surfaces. Most common nosocomial bacteria present and detected on inanimate hospital surfaces, using speciic marker genes, are presented in

<p>Bacterial Profiles and Antimicrobial Susceptibility Pattern of Isolates from Inanimate Hospital Environments at Tikur Anbessa Specialized Teaching Hospital, Addis Ababa, Ethiopia</p>

“…The hospital environment represents a new ecological place for medically important nosocomial pathogens, antibiotic-resistant microorganisms, and reservoirs of resistance genes, which have been common, found on various surfaces within hospitals (eg medical equipment, housekeeping surfaces, workplaces and lobby (furniture). 1 , 2 Studies investigating hospital environments reported that pathogens were ubiquitous in all hospital units but the interest was usually focused on intensive care and operation unit, especially due to the vulnerability of patients in these units. 3 There is also high antibiotic usage and invasive procedure from these units.…”

Bacterial profiles and their antimicrobial susceptibility pattern of Isolates from inanimate hospital environments at Tikur Anbessa Specialized Teaching Hospital, Addis Ababa, Ethiopia

“…Hospital environment represents a new ecological place for medically important nosocomial pathogens, antibiotic-resistant microorganisms and reservoirs of resistance gene, which have been commonly, found on various surfaces within hospitals (e.g. medical equipment, housekeeping surfaces, workplaces and lobby (furniture) [1,2]. Studies investigating hospital environments reported that pathogens were ubiquitous in all hospital units but the interest was usually focused on intensive care and operation unit, especially due to the vulnerability of patients in these units [3].…”

Infection Control in Dentistry and Drug-Resistant Infectious Agents: A Burning Issue. Part 2

“…The hazard for our reputation and insurance coverage is increasing with the possibility offered by molecular biology to identify dentally acquired infections [1]. Molecular biology and in vivo biosensors technology, to detect quorum sensing signaling molecules produced by airborne pathogenic bacteria, can prove the violations and noncompliances in dental settings and useful for accreditation surveys [43][44][45][46][47]58]. Nevertheless, antimicrobial surfaces and graphene-based antimicrobial nanomaterials seem to be promising to lower cross infection [122].…”

“…Fast and very sensitive molecular biological techniques (quantitative real-time polymerase chain reaction (PCR), multiplex PCR, microarray, next-generation sequencing technologies, etc.) and in vivo biosensors technology seem to be a very promising support to improve the knowledge on bioburden and biofouling, even due to not cultivable infectious agents by classical microbiological methods, and to monitor the effectiveness of item reprocessing [43][44][45][46][47].…”