PCRs that can be interpreted with gel electrophoresis alone through band size visualization.
Tests for Hepatitis will require viral load to be assessed though qPCR. I'd like to know if there are basic PCRs that can be important diagnostic tools for doctors.
I'd like to know as I'm interested in looking into the possibility of creating basic low cost PCR labs for small hospitals in rural areas of the the developing world.
Regular (aka "end-point") PCR is good at "presence, absence" tests. Like "does the patient have [whatever pathogen] present in their body?". It is not useful for telling the amount of the pathogen. Many labs instead rely on ELISA assays (for both detection and quantification).
PCR can sometimes be useful for detecting disease-associated alleles, IF there is a difference in allele sizes. But genetic mutations/disease-associated alleles are normally only diagnosed by DNA sequencing.
Do a literature search for the disease/pathogen you are interested in and what diagnostic tools already exist. Good luck!
HI Asif,
The qualitative HCV RNA tests use either a process called polymerase chain reaction (PCR) or a process called transcription-mediated amplification (TMA).
This type of qualitative test will report whether the hepatitis C virus is present in the bloodstream or not.
Regular (aka "end-point") PCR is good at "presence, absence" tests. Like "does the patient have [whatever pathogen] present in their body?". It is not useful for telling the amount of the pathogen. Many labs instead rely on ELISA assays (for both detection and quantification).
PCR can sometimes be useful for detecting disease-associated alleles, IF there is a difference in allele sizes. But genetic mutations/disease-associated alleles are normally only diagnosed by DNA sequencing.
Do a literature search for the disease/pathogen you are interested in and what diagnostic tools already exist. Good luck!
You can perform mutation detection using allele specific PCR for genes such as KRAS, BRAF, NRAS. They are usually done using real-time PCR now as it avoids PCR contamination, but all used to be done by running gels. You just need a different sized control reaction to make sure everything has worked. Also Cystic fibrosis mt testing to name but a few.
Dear Asif,
I would recommend you to make a literature search, as Katie recommended.
Second, you need to know which are the kits already used in your country or at least used in the region, which you will use in order to validate any kind of test you would like to develop. Read their manual of use or instructions in order to understand the composition of the kit, which reagents, controls and other components are used and their purpose, how the results looks like and how they are interpreted. Besides, it is the question of specificity and sensitivity evaluation, which is done on a certain number of patients, donors, other groups of people, as well as to be compared to a reference test (generally from big manufacturers).
Regarding the technique to be used, as Gillian mentioned, you MUST think about cross contamination and the ability / capacity of the laboratory staff to perform all the steps.
Testing of RAS genes is a harder task, as you must develop a sensible test to detect one possible mutation or one cell that contains the mutation from the others.
Good luck!!
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