Question 1: What are the advantages and disadvantages of Immunomagnetic Separation over other

phenotypic methods?
Answer: Immunomagnetic separation (IMS) is used to isolate target microorganisms from a mixed
bacterial culture using antigen-antibodies attraction in which magnetic beads are coated with
specific antibodies.
Advantages: 1. It is a highly specific method that helps to isolate rare microorganisms by using
specific antibodies that may be difficult to isolate using other phenotypic
methods.
2. It is a quick method; we can analyze the target organism in a few minutes or
hours while other methods require incubation for one or two days.
3. we can recover a high percentage of target cells as it gives accurate results and
can use these microorganisms for further analysis such as PCR or microscopy.
Disadvantages: 1. As IMS only depends on the antibodies to capture the microorganism but if
the antigen was not characterized on bacteria or if there is no antigen or very
low level of antigen then we will not able to work with the IMS method.
2. It does not provide information about antibody resistance, or virulence
factors where other genotype methods provide DNA sequence using PCR.
Also, in this antibodies can be easily replaced with inhibitors to bind the
target microorganism.

Question 2: What are positive and negative control in both IMS and LFA method. What purpose
do they serve in the experiment?
Answer: Immunomagnetic separation (IMS)
Positive control: The bacterial culture containing only Salmonella enterica inoculated in TSB
(Tryptic Soy Broth). The purpose of this control is to detect the antigen
present specifically on Salmonella enterica as the magnetic beads present in
tube are coated with Salmonella specific antibodies.
Negative Control: TSB (Tryptic Soy Broth) is used as a negative control to check the sterility of
the procedure.
Lateral Flow Assay (LFA):
Positive control: a Salmonella enterica dissolved in wash buffer is used as positive control
which makes a complex with colloidal gold conjugated antibodies and gives
two red lines on the Nitroso cellulose (NC) Membrane.
Negative control: a simple wash buffer with no bacterial colonies is used as a negative control.
The colloidal gold conjugated control indicator will give one red line
indicating the flow is working.