Advanced Molecular Techniques Of Detecting And Identifying Various Microbes (Essay Sample)

Molecular Microbiology
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Introduction
The new discoveries have been ongoing in an attempt to come up with advanced molecular techniques of detecting and identifying various microbes responsible for the various diseases more particularly the zoonotic diseases. Among the contributing factors to these advancements is the fact that these dangerous pathogens are frequently undergoing mutations thus making some of the techniques unsatisfactory and inconclusive while identifying the pathogens. This paper will discuss into details the potential benefits and setbacks of some molecular technologies in the identification of diseases spread between humans and animals by parasites, the pathogenic fungi, bacteria, and viruses (Dunn 2010, p. 16).
The (MLVA) Multiple-Locus Variable number tandem repeat Analysis is a molecular typing technique used to identify specific microorganisms (Barco, Barrucci, Olsen and Ricci 2013, p. 198). The procedure employs the natural variations in a tandem repeated in the DNA sequence found in different loci of the genes of different organisms. The technique is as well used for the assessment of bacterial molecular fingerprinting in the study of the transmission routes, to evaluate the causes of disease infection and the probable interventions such as the development of vaccinations and antibiotics against the zoonotic microbes (Keim et al. 2000, p. 2931).
The main benefits of MLVA that have impacted the public health sector significantly are that the technique is faster and safer in that it reduced the handling of live pathogenic bacteria when typing for these microorganisms (Graziani et al. 2008, p. 416). Similarly, the procedure considerably reduces the average typing time. MLVA as well provides a high resolution which is helpful in the determination of large and exceptionally complex outbreak analysis (Nadon et al. 2013, p 20565). The procedure is as well applicable in the large-scale automation by the use of state of the earth apparatus such as the automated sequencers fitted with the analytical software and the high-powered pipetting robots. The main disadvantages of this technique are that each organism requires a separate MLVA assay and the lack of the proper standardization for most of the published assays (Tiller et al. 2009, p. 2228).
The (MLST) Multi-locus sequence method of typing is a molecular procedure used in the characterization of bacterial isolates by use of the gene sequences usually the common seven house-keeping genes. The technique is centered on the principle that utilizes the multi-locus enzyme electrophoresis; however, the technology is different in that it allocates alleles within the multiple house-keeping loci through the process of DNA sequencing, instead of an indirect approach to electrophoretic mobility of the gene. The main benefits of this procedure are that the acquired profiles of the alleles can be easily related to the database that can be found on the internet that is contrasting to the typing methods which only compare the DNA fragment on the gel (Sabat, et al. 2013, p. 2038).
Similarly, the gene profiles comparison can as well be attained from the clinical material of the PCR amplification directly from either CSF or blood; therefore, the isolates can be characterized even if the pathogen strains in question cannot be isolated. The limitations of this technique are that even though the method seems to be the best, it is very expensive to perform. Similarly, MLST occasionally lacks the power to distinguish the various bacterial strains, and this limits its practical use in the epidemiological examinations. However, to improve on the power discrimination of different isolates when using MLST, a new technique known as Multi-virulence-locus sequence typing has been established by use of Listeria mono-cytogenetic. The MVLST increases the advantages of MLST. However, the method only targets the virulence genes of which the majorities are more polymorphic rather than the housekeeping genes (Gilmour et al. 2013, p. 5).
The (WGS) whole genome sequencing equally recognized as the complete genome sequencing procedure is a technique used to establish an entire DNA sequence in both human beings as well as the genomes of the pathogenic micro-organism. This process involves sequencing all chromosomal DNA of an organism and the DNA within the mitochondria. The Benefits of these techniques is that it can be possible to create a personalized plan and treating a zoonotic disease is possible basing on both the mutant genes causing the disease and the patient’s gene. The benefits of this approach are that it is feasible to understand the misrelated genes through genotyping the cancer cells thus selecting the best medication which exposes the patient to less toxic chemotherapy. Similarly, through (WGS) the unknown genes can be fully recognized contrary to the traditional molecular techniques which examine the trouble maker genes (Brinkman et al. 2013, p. 566).
Lastly, gene predisposition due to the environmental variations can be without difficulty identified and moderated. The potential disadvantages of (GWS) are that the roles of a majority of the genes in the human genome are unknown with little or minimal understanding. Thus, most of the information obtained by studying the human genome is impracticable. Similarly, most of the technicians are not proficient in the interpretation of genomic data because the amount of information in the genome is too much and the measures of maintaining the safety of this genomic information are still new. Besides, the patient’s genome might have information that is unnecessary to the physicians, for instance, the patient whose genome sequencing is performed to establish the effective treatment for the high amount of cholesterol. The researchers may ascertain an entirely unrelated allele of a terminal disease without an effective treatment (Brinkman et al. 2013, p. 569).
The (PFGE) Pulsed-field gel electrophoresis technique is a method used by researchers to come up with a DNA fingerprint for the zoonotic a bacterial isolate (Noller et al. 2003, p. 678). The main advantages of this approach are that it can be utilized as a general generic subtyping process for various with the restriction of the enzyme as well as electrophoresis optimization of each bacterial species. The technique as well is capable of producing stable DNA restriction patterns which can be reproduced. Moreover, the PFGE method has a high discriminating power as compared to other methods like the ribotyping or the multi-locus sequence typing.
Despite the outstanding advantages, the technique as well has its setbacks; first to carry out the whole procedure of PFGE is time-consuming and not all isolates especially those which are unrelated can be discriminated by this procedure (MacCannell 2013, p. 631). Besides, the DNA restriction of the isolates patterns in question do at times vary between the technicians, the PFGE as well does not optimize the separation of every part in the gel instantaneously. In PFGE, the bands that contain the same size might not be able to come from one point of the chromosome of the pathogen that is being investigated. The other limitations of this technique are that this approach cannot establish some pathogen strains. Finally, this method cannot differentiate the isolated under investigation to the same degree (Gui and Patel 2011, p. 1).
Pulse Net is a network currently made of 83 international public health laboratories as well as those of food regulatory. This network brings together people from different parts of the globe more particularly those who have been exposed to pathogenic microbes or those who ate contaminated food. The benefits of Pulse Net are that they help the researchers to determine places where the food safety procedures have failed and developed appropriate solutions that can stop future illnesses. Similarly, Pulse Net has in a great deal helped the scientists to determine if a disease outbreak is or will occurring, the network greatly allows the state departments to identify disease outbreaks and the causative agents in a very short period (Swaminathan et al. 2006, p 39).
Besides, the epidemiologists across the globe utilize the information from Pulse Net to help in determine what exactly is causing sickness in people. The organization as well works with the public health governing agencies, like (FDA) Food and Drug Administration in the U.S and the Agricultural Department of U.S Food Safety. The Inspection Service (USDA/FSIS), in establishing solutions to terminate zoonotic disease outbreaks and coming up with the way of averting the situation (Gerner-Smidt et al. 2006, p. 16). However, Pulse Net is facing many challenges, first, due to limited funds and the shrinking of the state budgets; the Pulse Net activities have significantly been affected. Similarly, the insufficient staffing in a...