CLS 513 Advanced Diagnostic Microbiology

Question: Research Article Rapid and safe one-step extraction method for the identification of Brucella strains at genus and species level by MALDI-TOF mass spectrometry. You should give a brief introduction and methodology to the paper, Then you talk about the findings and the meaning of each finding. Answer: Brucellosis, a zoonotic disease, is still consider to be a threat to the domestic livestock around the world and remains a concern for the livestock industries due to financial losses and public health. Being a zoonotic disease, it can infect human through contaminated dairy products or meat which leads to the need of accurate and rapid identification method for Brucellosis. Currents method for identification are time consuming and expensive and recent development in MALDI-TOF base mass spectrometry has emerged as a viable replacement. This article have designed a new protein extraction protocol along with a generation of new customized reference database. The database contained the list of wide spread species of Brucella strains. Then, this database were run against the infected sample collected from infected livestock. The assay was able identify 99.5 per cent of Brucella strains at genus level and 97 per cent of Brucella strains at species level. This signifies that the microbial isolation at differing culture conditions did not affect the assays. Furthermore, reliability of this assay was confirmed by the fact that it did not have any adverse effect on the protein stability while stored and sent to reference databases for comparison. However, this MALDI-TOF assay was not able to deduce accurate phylogenetic tree signifying that protein profile does not correspond with evolution of genes. To summarize, based on the accumulated data, this MALDI-TOF assay is a reliable, accurate and fast method for the identification of Brucella strains. After thorough revision and study of this article, following three questions have been raised: What kind of steps and factors are necessary and should be considered to upscale their protocol, so that it can be used industry and health sector immediately? What measures can be taken or incorporated in their assay protocol, so that it can accurately decipher a dendrogram? In countries like Africa, time taken to ship a sample from remote area to a lab might take more than 72 hours. Would this assay will still be effective and reliable if the samples were stored more than 72 hours?

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CLS 513 Advanced Diagnostic Microbiology

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