Enhanced chemiluminescent (nonisotopic) detection of rice tungro bacilliform virus DNA.

Date

10-1996

Degree

Bachelor of Science in Biology

Major Course

Major in Cell Biology

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Merlyn S. Mendioro

Abstract

A study was conducted to evolve a system that would allow early detection of the rice tungro bacilliform virus (RTBV) at DNA level in infected plants. The study involves: 1) generation of probes; 2) establishment of the assay (tissue prints); 3) optimization of the ECL (enhanced chemiluminescence) direct nucleic acid labeling and detection systems with tissue prints as assay; and 4) to test whether the probe is general or specific to the four RTBV variants/isolates (L, GI, G2, and Ic). Only two possible probes (clones 2a and 5d) were obtained out of the 20 bacterial colonies screened. Between 2a and 5d, the latter was more consistent in detecting the RTBV DNA. Membranes which were treated with the denaturation solution prior to prehybridization and washed at 52°C with 0.1X SSC without SDS (stringency) gave the best results. The healthy samples can be readily differentiated from the infected ones. The B/S-infected plant was consistent in generating the most intense signal and the virus therefore occurs at a higher titer in doubly-infected plants.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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