Describe what is meant by phenotypic, genotypic and immunological test methods o
ID: 181081 • Letter: D
Question
Describe what is meant by phenotypic, genotypic and immunological test methods of identification of microbe.
What is the significance of doing an ELISA or Western Blot test
What are some ways we visualize the antigen/antibody interaction in the lab List some advantages and disadvantages of the Salk and Sabin vaccine Why would a polymerase chain reaction be done?
What is gel electrophoresis and why is it done?
Be familiar with spotlighted microbes discussed in lecture.
High light •Part of normal flora, found in intestine, vagina.•Can inhibit other bacteria, yeasts –ferments carbohydrates into lactic acid•improves bowel
Explanation / Answer
Ques-1: Describe what is meant by phenotypic, genotypic and immunological test methods of identification of microbe.
Gram staining is the phenotypic study is useful to study the expression of the type of cell wall material in microbes for example, bacteria
Genotypic identification of microbe:
The genetic analysis of protein expression as per gene can be specific for specific microbe therefore; DNA microarray analysis of gene and their mRNA components followed by complementary DNA synthesis is useful to unravel the genome of bacteria. DNA recombination techniques are also useful to unravel the mechanism of genes involved in the microbe
Immunological study
The phenotypic method of immunological test is by studying the expression of protein on the cell membranes or inside the cell (for example lipopolysaccharides) using primary and secondary antibodies using immunohistochemistry. Double immunolabelling is the methodology that is used in indirect ELISA. Initially wash the sample contain antigen with a primary antibody followed by the elimination of the unbound antigen. Now apply enzyme linked secondary antibodies to the primary antibodies where secondary antibody binds to the primary antibody’s nonspecific Fc region followed by the washing procedure to eliminate any unbound
Antibody-enzyme conjugates. Now any fluorescent indicator is added to produce an electrochemical signal.
Ques-2: What is the significance of doing an ELISA or Western Blot test
The ELISA (Enzyme Linked Immuno-Sorbent Assay) is used to detect the presence of antibody or an antigen in the test sample. In this technique, two antibodies are used of which one is coupled to an enzyme (so the name “enzyme linked” and this allows the fluorogenic substrate to generate a signal) and another is specific to the antigen. This technique can be used to detect the presence of both antigen and antibodies such as those of HIV etc.
Western blotting or protein immunoblotting technique is an analytical technique used to identify specific proteins & their interactions with other partners in a sample, the sample may be tissue homogenate or extract.
Tests need to be performed to analyze the neuropathology of the hippocampus and cortex by western blot analysis and also by immunohistochemistry for the localized expression to identify Down syndrome over expression protein at both cellular and receptor level