Greetings. I would be grateful if you could interpret below Elisa report about a broiler breeder 46 week vaccinated on date 12/02/2022 with Ma5 and clone 30 spray. This farm is composed of 5 pens. I would like to know:
1- How can I specify validity of internal controls of the Elisa report. Is it necessary to be similar to that of certificate of analysis? If no what is the difference between them to be accept or shall I depend on the reader to validate it.
2- All the pens have the same mean titer and zero %CV although the mean titer within the expected range.
3- Why does that phenomena occur and what is the corrective action? Thanks and regards
Elisa report Mean titer 18937 Titer range reference control cr100 RF16 6000-12000 Min-max titer 18937-18937 Mean titer Ref Control 6983 GMT 18937 Internal controls .199 and .241 for negative %CV 0 .854 and .908 for positive control NO of samples 18 Raw OD for all samples 4.000 Neg/su/pos 0/0/18 Titer Group 11 Result positive
First of all mention Is it ELISA of ND or IB? also mention name of Kit used for ELISA?
Then mention no of vaccine shots done from day one to 46 weeks of age, for which ELISA is required. After that CV%age zero for 18 samples doesn't look possible, as same value for 18 samples is not possible practically.
Respected Sir
Deeply sorry I tried to attach Elisa report but I didn't know how to attach it any how
Biochek Elisa
ND test
No of samples tested 18
No of vaccine shot 12from day to 20 weeks after that one shot monthly up to 46weeks
Respected Dr. Mohammed
The ND test followed IB test with the same samples . Elisa reader did not give any sign to indicate the test is in valid when the result was saved .This phenomena occur before with IBD. All the samples except the internal and external control the raw OD was 4.000 . for that reason I asked how could I specify the validity of internal controls . Should they must be the same as that in certificate of analysis. Definitely no .shall I depend on the the reader to specify it .
Hi,
As you are using BioChek ELISA, you can find the interpretation guideline in the below link
https://www.biochek.com/wp-content/uploads/2018/01/BIOCHEK-INTERPRETATION-Manual.pdf?x14571
Or you can approach your Kit supplier to get technical support from the BioCheck team.
ZERO CV% is quite doubtful as it never happens. As your MIN-MAX are shown same titer, CV came up with ZERO. If possible please share the Individual sample titer. If all 18 samples are showing same titers then there could be some error in the protocol or reading.
Respected Sir if there is something wrong in protocol or reading that may be indicated by the reader. How can that happen since result was saved as usual?
Hi Salah Mohammed
in my experience with ELISA reader and software (irrespective of any brand), reader will only indicate if there is any deviations in controls (Pos & Neg). for the samples, it will show as it read.
in your report, all the samples are showing same titers, that is quite impossible. either you need to re test the samples and check whether same problem persist or need to check the reader for calibration or other possible error with the reader supplier.
I.B Elisa titers 18000 indicate exposure to disease. Vaccine protective titers are around 7000. Coefficient of variation (C.V%) is never negative and anything below 20% is considered good. The results should be rendered back to the concerned lab. For clarity. Test again elsewhere.
In general good practice to start your Elisa testing as
1- make sure your washing system is clean primer the wash machine before running the test and check the water PH should be distill water no salt residue
2- use through for each conjugate separately measuring actual amount needed for the test never return any amount left to the original bottle to avoid cross contamination
3- bring all kits reagents to the room temperature a lest 2 hours
4-multichannel pipette should be maintained in a good condition and cleaned regularly. Use tips that fit with your multichannel pipette
5-Regularly preform Reader testing for ODs value with calibration micro titer plate
6-a good practice to share your results with technical services provider of the Elisa kits to evaluate any deviation of the results it is very difficult to conclude what is wrong we need to see your previous results and the expected value of kit certificate of analysis
Hafiz Babiker Hello Sir
Thank u so much for comment and valuable advice. I would be grateful if you provide me your Email to communicate with you
Thanks and cheers
Dr. SalahSldean Ali
Saudi Arabia
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