Immunocapture Polymerase Chain Reaction (IC-PCR) and Nucleic Acid Hybridzation Techniques for Detection of Spiroplasma citri
Immunocapture Polymerase Chain Reaction (IC-PCR) and Nucleic Acid Hybridzation Techniques for Detection of Spiroplasma citri
Om-Hashem M. El Banaa l, A.A. Abou Zeid2, Fawzia I. Moursy3 and Azza, G. Farag
ABSTRACT
Spiroplasma citri Saglio et al is the type species of the genus spiroplasma. Spiralin gene is the gene responsible for spiralin which is the major protein of Spiroplasma citri (S. citri Saglio et al), the causal organism of citrus stubborn disease. RecA gene protein is the enzyme primarily responsible for the homologous recombination of chromosomal DNA in bacteria. The Immunocapture- Polymerase Chain Reaction (IC-PCR) technique was applied for detection of S. citri Saglio et al in which the Spiroplasma was captured with the specific polyclonal antibodies on a solid-phase. Specific primers based on the sequence of the Morocco strains of S. citri Saglio (R8A2HP and G113) were used. Two pairs of primers SC. SC’ and SC8. SC9 were used. PCR fragment of correct size 330bp was amplified with primers SC. SC expressing spiralin gene and 760bp was amplified with primers SC8, SC9 for recA gene. No amplified products were obtained from samples of healthy citrus trees. Nucleic acid hybridization using non-radioactive DNA probes through southern blot hybridization technique revealed the sensitivity of this technique for detection of S. citri Saglio et al infected citrus trees.
To share on other social networks, click on any share button. What are these?
