engleski

Single-Strand Conformation Polymorphism Analysis for Differentiating Phytoplasma Strains

Single-strand conformation polymorphism (SSCP) analysis is a sensitive and rapid technique for detecting DNA polymorphism and mutations in PCR amplified fragments. Due to its technical simplicity, it is widely used as a screening tool in various investigations, ranging from clinical diagnosis of human hereditary diseases to the characterization of microbial communities. It was shown that this method can also be successfully used as a tool for detection of molecular variability in phytoplasma evolutionary conserved house-keeping genes such as 16S rRNA and tuf as well as more variable genes, revealing the presence of polymorphism undetected by routine RFLP analyses. Reliability of SSCP was confirmed by the multiple alignments and phylogenetic analyses of representative sequences showing different SSCP profiles. However, it is not yet broadly applied in phytoplasma research. This technique offers inexpensive, convenient and sensitive method for determining sequence variation and differentiation of phytoplasma strains which can particularly be suitable in epidemiological studies or as a fast screening typing tool when dealing with a large number of field samples.

methods, microbiology, phytoplasma, sscp, variability

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