JOURNAL ARTICLE REVIEW (SCIENCE) 1 Journal Article Review Bacterial identification relies primarily on culture based techniques which need around 12hoursfortheisolationandmorethan48hoursfor speciesevaluationand identification. The matrix assisted lased desorption is one of the best technique which is used for the identification of bacterial species. This essay goal is to critique a journal article and evaluate the benefits and drawbacks of both molecular and phenotypic techniques. This essay will emphasis the PCR, and MALDI-TOF molecular methods and routine phenotypic methods and analysis the fundamental concept of these techniques in order to identify the bacterial species. The title of journal paper is Comparison of Two Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Methods with Conventional Phenotypic Identification for Routine Identification of Bacteria to the Species Level which is based on the identification of bacteria. It was written by Abdessalam Cherkaoui, Jonathan Hibbs, Ste´phane Emonet, Manuela Tangomo, Myriam Girard, Patrice Francois, and Jacques Schrenzel. According to this journal paper, the identification of bacteria by using pathogen is more crucial and complex. To reduce such kind of issues the clinical laboratories developed two methods that are more efficient such as molecular method and phenotypic method (Cherkaoui, et al., 2010). Molecular is one of the advanced methods that have the potential to identify the rapid bacterial and it is a more effective technique rather than other. The PCR is a part of molecular technique which is more sensitive in order to identify the bacteria and in current clinical this technique is used to evaluate the bacterial spices. The researcher also observed that the quality related issues in this method are very less and PCR based evaluations are further complicated through required getting the sample ready. Many PCR based methods are performed on the routine clinical labs and they need nucleic acids gathered from the isolated colonies. It is analysed that the direct PCR method is used to detect a small number of species. According to this research paper, the matrix assisted lased desorption ionization time of flight mass spectrometry (MALDI TOF MS) is additional kind of molecular method that is supportive for the diagnostic of bacteria. The main advantage of this technique is that
JOURNAL ARTICLE REVIEW (SCIENCE) 2 it is used as a technique for analysing the applied microbiology. Researcher argued that this technique also indicates rapid turnaround time, modest reagent costs and the low sample volume. It also helps for producing the pathognomonic pattern which provides complete information about bacterial species. The main disadvantage of this method is that it has very less short turnaround time due to which it is not much popular as compared to the PCR technique. It has considered as the ribosomal from Escherichia coli and researcher analysed the use of MALDI-TOF MS technique for evaluation and documentation of microorganisms based on the ribosomal. A current study about MALDI-TOF MS evaluated that this technique is more useful for analysing the bacterial species. This journal paper reduced the research gapes faced by the previous study and improved the databases and analytical tools for better understanding. According to the author, the MALDI-TOF MS technique is usefulness for the testing of clinical and it is very important to identify the applied method is appropriated or not. The major purpose of this paper is to assess the performance and effectiveness of the MALDI-TOF MS under real routine laboratory situation. The other advantage of this technique is that it has the potential to identify most isolates process in less time with better outcomes only if the MALDI-TOF MS has been failed. Moreover, the researcher also suggested that MALDI-TOF MS method is also valuable for determining burkholderia cepacia from cystic fibrosis individuals. This journal paper compared two major methods such as MALDI-TOF MS and routinely based method for the documentation of bacterial species. In which the researcher used PCR and sequence based method in order to solve discrepancies and identified the accuracy of speciation, cost and turnaround time of both these techniques. In the routine phenotypic method, the isolates are evaluated based on the plate morphologies and it takes around 24 hours after the specimen is collected. Such kind of method is more relevant to the scientific microbiology meanwhile many outcomes can be gathered 1 era earlier and it takes more time as compared to the molecular method. The researcher also identified that the phenotypic method takes more than 24 hours for completingtheidentification test.Thecostofphenotypicbacterialidentification method is around US$10 per isolate which is more expansive as compared to the PCR technique.
JOURNAL ARTICLE REVIEW (SCIENCE) 3 The main advantage of the molecular technique is that it has the ability to evaluate and identify the multiple organisms with sufficient specificity and sensitivity. Moreover, this kind of technique is more efficient and accurate in order to identify the bacterial species but it required more time for identifying the bacterial species. With the help of this technique, individuals can determine the rapid bacterial and PCR is a more sensitive part of the molecular method. According to the researcher, the molecular technique can be used as an analytic tool that may prove useful diagnostically. Researcher argued that the major disadvantage of the molecular method is that it depends on the laboratory accreditation requirement and testing methods due to which it cannot be used to perform molecular testing. It is observed that it need more space in the clinic laboratory which creates a problem for a small laboratory. The other drawback of this technique is that it is more costly as compared to the other methods and it also required more knowledge during the identification of bacterial species. The key advantage of phenotypic identification method identified by the author is that it is very easy to understand apply rather than molecular method. With the help of this technique, consumers can evaluate the bacterial species and in terms of equipment costs it less expansive. The researcher analysed that such kind of method also provides extra information for the evaluation of the bacterium. According to the researcher, the phenotypic identification method is not much popular because it cannot be used for the identification of multiple bacterial species. Moreover, it is less efficient as compared to the molecular technique. From this journal paper, it is observed that the phenotypic identification method is more complex and slow in order to identify the bacterial species. The researcher analysed that the speed of the assays is very high in the case of MALDI- TOF MS but it is very low in the case of phenotypic identification technique. It is also estimated that the turnaround times of MALDI-TOF MS and molecular method is very short due to which MALDI-TOF MS is not much popular technique. As compared with the phenotypic technique the overall turnaround time is very high due to which most clinical laboratories use this type of technique for identification of bacterial species.
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JOURNAL ARTICLE REVIEW (SCIENCE) 4 From this study, it is analysed that the molecular identification technique has the capability to detect the multiple organisms in a single assay with more specificity and sensitivity but the phenotypic method does not identify multiple organisms and it has less sensitivity and specificity. The author also identified that the cost reduction, technical training and quality requirements and less turnaround time make MALDI-TOF MS method more quite as compared to the phenotypic method. The labour intensive of deploying the equipment are very less in the case of the molecular method but in the case of phenotype method, this intensive is very high. From this paper, it is found that the cost implication of phenotypic technique is around US$10 per isolate but in the case of molecular based identification technique is cost does not exceed $0.50. There are few limitations of phenotypic assays such as it is not much efficient, very slow in operation, and less accuracy in the identification of bacterial species. It is concluded that MALDI-TOF MS molecular is one of the best technique in order to identify the bacterial species. From this investigation, it is observed that the advanced MALDI-TOF MS technique is moreprecise and efficient for the clinical laboratory. The MALDI-TOF MS technique main limitation is that it has a short turnaround time and less readily interpretable data due to which it is not much popular for protein evaluation and determination. It has resulted that the large confidence identification with the help of MALDI-TOF MS technique had better accuracy and huge confidence identification through Shimadzu MS and Bruker MS were 99% accurate as compared with the conventional technique. This essay explained the two types of methods that are a molecular and phenotypic technique for identification of bacterial species. It is concluded that the molecular technique can be used to identify multiple organisms with better sensitivity and specificity. Thisessay also evaluated that advantages and disadvantages of both molecular and phenotypic techniques and a comparison section are also added for better understanding. Moreover, the types of molecular identification technique and routine phenotypic methods are also explained with their characteristics.
JOURNAL ARTICLE REVIEW (SCIENCE) 5 References Cherkaoui, A., Hibbs, J., Emonet, S., Tangomo, M., Girard, M., Francois, P. and Schrenzel, J., ( 2010) Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry methods with conventional phenotypic identification for routine identification of bacteria to the species level.Journal of clinical microbiology,48(4), pp.1169-1175.