Small Subunit Ribosomal DNA-based Variation Analysis And Detection Of Plasmodium Vivax Of China

Malaria is one of the most serious parasitic diseases especially in the tropical and sub-tropical regions of the world. The prevention and treatment of malaria has being highly attached importance in the domain of the medical sciences. The structure of ribo-somal RNA gene units is identical in most eukuryotes of which partial sequence is highly conser(?)ative and homologous amongst the different species of eukuryotes, but partial sequence is variable even in the same species of eukuryote because the rate of nucleotide transitions, transversions and deletions or insertions throughout evolution is not constant. It is demonstrated that rDNA (rRNA) can be used not only as significant foundation of parasitic classification based on sequence variation analysis, but also as ideal target based on special sequence for detection of parasites in the host. Up till now, it is not found that the articles on intra-species variation analysis based on rDNA sequences of Plasmodium and detection based on SSUrDNA for diagnosis of vivax malaria.In this paper, according to known SSUrDNA sequences of Plasmodium vivax, Plasmodium falciparum, Plasmodium malariae, Plasmodium berghei, Plasmodium lo-phurae, Trypanosoma cruzi, Leishmania donovani, Sarcocystis muris, Pneumocystis carinii and Human being, the sequences of oligonucleotide primer pairs specific to Plasmodium and to P. vivax, which incorporated restriction endonucleases Hinc II and Pst I, were defined respectively with computer programming. The desired DNA fragment size of P. vivax, about 640 base pairs in the length, were directly amplified by two temperature point polymerase chain reaction from conventionally extracted genomic DNA of blood samples of P. vivax malaria patients from Anhui, Fujian, Hainan, Sichuan and Yunnan provinces of China (two cases/each province), but none of cultivated erythro-cytic stage of P. falciparum isolates (FCC/AH, FCC1/HN and FCC/YN), promastig-ote of L. donovani, tachyzoite of T. gondii and Human. The SSUrDNA fragments amplified of P. vivax were identified with methods of digestion of restriction endonucleases and Northern blot hybridization.The SSUrDNA fragments amplified of P. vivax isolates from five provinces were digested with Hinc II/Pst I and ligated to vector (M13mpl8 and M13mp19) predigested with the same restriction endonucleases. At least two recombinants inserted with the...