Bacteriophage based bacterial detection | Phage | Phage Detection | Basic Science Series
• A bacteriophage is a virus that infects and replicates within bacteria. They are composed of proteins that have DNA or RNA as their genetic material. it has a small neck section that connects tail sheath with head. To the tail end, it contains tail fibers that are required for binding with bacterial receptors.
• Bacteriophages attack specific bacterial populations. This specificity means a bacteriophage can infect only certain bacteria bearing receptors to which they can bind, which in turn, determines the phage's host range. Attachment of phage with bacteria surface receptor is called phage docking. Host growth conditions also influence the ability of the phage to attach and invade them. Staphylococcus phage will attack only staph. Ecoli phages will attack specified strains of Ecoli. Therefore they are very full tools. Now please see chapter-3 understand the mechanism of bacteriophage mediated lysis.
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• There are two types of phages lytic and lysogenic phages. Specifically lytic phages are the phages that are useful since they multiply in the host and eventually lyse the cell.
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• The lysogenic phages incorporate their DNA into bacteria cells and remain latent. Their chromosome becomes integrated into a specific section of the host cell chromosome. We are specifically focusing on lytic phages in this presentation.
• Antimicrobial resistance is the ability of a microbe to resist the effects of antibiotics. Resistant microbes are more difficult to treat, requiring alternative medications or higher doses of antimicrobial agents. In my previous episodes Chapter- 15 I have explained in detail about antibiotic resistance and antibiotics.
• In the previous presentation, Chapter-20 I have explained the method of detection of bacteria using the specific examples of MRSA. The methods mentioned in the presentation require more than a day. Therefore advanced methods are required. On of the method to detect is bacteriophage-based detection of antibiotic-resistant bacteria. I will explain this by taking the example of MRSA in following steps-
• Step one: Bacterial sample preparation: For phage replication bacteria need to be in a specific number so in the low number you need to incubate in the media to increase the number or if you have a high amount then you need to dilute the bacteria. The preliminary quantification can be done by using UV spectrophotometry. To understand how UV spectrophotometry works please refer chapter- 14.
• Step-2: Now at a particular concentration of bacteria. Methicillin is added to the culture. Other antibiotics can be tested separately. Now if bacteria is sensitive bacteria will not grow. If bacteria is resistant then the bacteria will grow. Now you want to check whether the bacteria survived.
Step-3: You will add staph phage which should multiply after hitting the bacteria and the phage population will go higher. Phage is proteins and you can use any method of detection to find out whether there is a significant increase in protein content. Method of phage detection includes antibody-based methods like ELISA, mass spectrometry, nuclear magnetic resonance.
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