EVALUATION OF SENSTITIVITY OF PCR AMPLIFICATION FOR ADENOVIRUS AND PORCINE PARVOVIURS DETECTION BY THREE DIFFERENT METHODS / 微生物学通报

ABSTRACT

Using specimens spiked with adenovirus or porcine parvovirus, several DNA extraction methods were evaluated for their ability to remove polymerase chain reaction (PCR) inhibitors from stool samples. It was found that PCR inhibition could be partially overcome by extracting viral DNA with guanidine thiocyanate ( GuScN) or chelex resin methods, but PCR inhibition could be hardly overcome by extracting viral DNA with traditional proteinase-K phenol chloroform extraction method. For adenoviurs (dsDNA) in stool samples, GuScN, chelex resin, proteinase-K phenol chloroform methods respectively allowed 5 TCID50, 50TCID50, 50 TCID50 titer to be detected. For porcine parvovirus (ssD-NA) in stool samples, three protocols respectively allowed 10 TCID50, 50 TCD50, 100 TCID50 to be detected. . Their rapidity and low cost make GuScN and chelex resin extraction methods the suitable for routine diagnostic testing.