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Qualitative Elisa Test for Detection of Bt Protein in Brinjal Leaf Tissue Samples

M. Mahmuda Khatun
Senior Scientific Officer
Biotechnology Division, BARI, Gazipur

D. Khanam
Principle Scientific Officer
Biotechnology Division, BARI, Gazipur

M. Al-Amin
Chief Scientific Officer
Biotechnology Division, BARI, Gazipur

Abstract/ Executive Summary:

Nine Bt varieties were tested to determine the presence and absence of Bt protein in brinjal leaf tissue samples. A total of three thousand one hundred ninety samples were tested where two thousand nine hundred forty samples showed Bt positive. Among the varieties tested, Khatkhatia, Singhnath and Kazla showed 99% Bt positive samples. Dohazari and ISD 006 showed 98% Bt positive samples. Uttara and Nayantara showed 97% while Islampuri showed 74% Bt positive samples and Chaga showed only 20% Bt positive samples.

Keywords: Bt protein, Brinjal, Qualitative ELISA

Location: Biotechnology Division, BARI, Gazipur

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Brinjal

Objectives:

To detect Bt protein in brinjal leaf tissue samples

Methodology:

Procedure:

Anti-cry 1Ac conjugate (50 µl/well) was added in pre-coated plate.
Sample extract, positive control and negative control (50 µl) were added to the appropriate wells. 1X sample extraction buffer was also added in blank well. The plate was incubated at room temperature for 45 minutes.
The plates were washed four times with 1X Buffer A (wash buffer). Then the plates were patted dry on blotting paper to remove excess buffer.
Substrate (100 µl) was added per well.
The plate was incubated at room temperature for 15 min.
After 15 min. 100 µl stop solution was added per well.
Data interpretation was made by visual observation. Yellow color indicated Bt positive samples.

Source of Information: Biotechnology Division (BARI) Annual Report, 2012-2013,p. 69, Gazipur

Article Link (Online Journal):

Funding Source:

1. United States Agency for International Development

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Nine Bt varieties were tested to determine the presence and absence of Bt protein in brinjal leaf tissue samples. Result indicated that in Kazla, 468 sample showed Bt positive out of 470 samples. In Singhnath, 310 sample showed Bt positive out of 311 samples. In Khatkhatia, 320 sample showed Bt positive out of 321 samples. On the other hand, in Dohazari, 314 sample showed Bt positive out of 320 samples and in ISD 006, 338 sample showed Bt positive out of 344 samples. In case of Uttara, 460 sample showed Bt positive out of 470 samples and in Nayantara, 300 sample showed Bt positive out of 308 samples while in Islampuri, 428 sample showed Bt positive out of 576 samples and Chaga showed 2 Bt positive out of 10 samples.

Knowledge Management: Report/Proceedings