Establishment of a Bioinformatics Method for Detecting RNA Viral Pathogen Causing Unknown Infectious Diseases

摘要

In this study, HIV was taken as the study object to establish a method for the detection of RNA viral pathogen causing unknown infectious disease. The serum from patients with acquired immune deficiency syndrome (AIDS) were filtered and treated with DNase to extract endogenous DNA using DNase-sequence-independent single primer amplification (SISP). Viral RNA from the serum was extracted, reverse transcripted, and the double-stranded cDNA was synthesized. The double-stranded cDNA was digested with TaQ and added to connect the adapter molecule, the pathogenic gene were amplified nonspecifically with the adapter molecule as primers, then all the PCR products were sequenced and analyzed. The results showed that a number of exogenous DNA fragments were obtained using DNase-sequence-independent single primer amplification. The result that All PCR products were sequenced and analyzed verified that all sequences were consistent with those of the known pathogenic genes (HIV virus). In conclusion, a viral RNA pathogen can be determined using DNase-sequence-independent single primer amplification.