การตรวจหาชนิดของพิษงูกะปะ Calloselasma rhodostoma และพิษงูเห่า Naja kaouthia ด้วยวิธีดอต-อิไลซา
Treatment of snake-bite using species-specific antivenoms requires identification of the snake venom. One method to achieve this is to use ELISA (enzyme-linked immunosorbent assay) technique. However, this method has some disadvantages including long incubation time and the requirement of expensive...
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| Format: | Theses and Dissertations |
| Language: | Thai |
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จุฬาลงกรณ์มหาวิทยาลัย
2002
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| Online Access: | https://digiverse.chula.ac.th/Info/item/dc:35767 |
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| Summary: | Treatment of snake-bite using species-specific antivenoms requires identification of the snake venom. One method to achieve this is to use ELISA (enzyme-linked immunosorbent assay) technique. However, this method has some disadvantages including long incubation time and the requirement of expensive instruments. The objective of this study is to develop a practical method for detection of snake venom by dot-ELISA on nitrocellulose paper using Calloselasma rhodostoma and Naja kaouthia venoms as models. Several parameters including purification method of anti-venom lgG, time for incubation and washing, volume and concentration of reagents and substrate for enzymatic detection were optimized. The sensitivity of the detection was 5 mu g/mL for Calloselasma rhodostoma and 1 mu g/mL for Naja kaouthia. No cross-reaction between the two snake venoms was observed between 0.2-25 mu g/mL. For comparison, the same ELISA were performed in microtiter plates and showed detection limit of 1 ng/mL for Calloselasma rhodostoma and Naja kaouthia venoms. In spite of poorer sensitivity, the developed dot-ELISA technique is much less time consuming and requires no special instrument which might be advantageous for field-test. |
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