Isolation of DNA from a single helminth using new developed kit in Iran and its PCR analysis

ABSTRACT

Nematodes are among the most common and important parasites of man and animal. DNA of a single worm can be used for several purposes, such as identification to the species, subspecies, strain and antihelmenthic resistance. DNA extraction from a single small worm using traditional methods such as phenol extraction technique faces serious problems. DNA from 20 single Haemonchus contortus was isolated using DNA isolation kit newly designed in Iran by the Research Unit of Molecular Biological System Transfer [MBST] based on the specific binding of DNA to the carrier. The genomic DNA was amplified using specific primers derived from beta-tubulin isotype 1 in PCR. The specificity of the PCR products was determined using semi-nested PCR technique. Specific PCR-product from beta-tubulin gene could be amplified with 1 ng, 100 pg and 10 pg DNA. The used DNA extraction method was safe, with high quality and quantity, fast, easy to handle and not costly for genetic analysis of even a single small worm. The Iran produced DNA extraction Kit is grounded on a selective binding of nucleic acids to a silica-based membrane and is recommended for the isolation of DNA from even small amount of biological materials