1b and 2a HCV subtypes we re mainly reported in ChinaHerein ,a 3b subtype strain of HCV was fo und with gene chip methodFirstly, the 5’ NCR(non-coding region)was s elected as target sequence,and 171 related seque...1b and 2a HCV subtypes we re mainly reported in ChinaHerein ,a 3b subtype strain of HCV was fo und with gene chip methodFirstly, the 5’ NCR(non-coding region)was s elected as target sequence,and 171 related sequences in GeneBank wer e alignedAccording to the alignme nt results,eleven DNA probes were acquired and oligonucleotide array s were constructed for distinguish ing the HCV main genotypes and sub types,which did not contain specif ic probes of genotypes 4 and 5,but genotype 1, subtypes 1a/1c and 1b, genotype 2,subtypes 2a/2c and 2b,g enotype 3,subtypes 3a and 3b and g enotype 6,subtype 6aSecondly,foll owing reverse transcription of HCV RNA,nested-PCR was applied to harv est the 260 bp target DNA fragment approximatelyAfter specific ampli fication of target DNA and array h ybridization,the solid phase alkal ine phosphatase reaction indicated visually the results that the DNA fragment contains specific sequenc e of genotype 3 and subtype 3bBy sequencing the target DNA fragment ,and analyzing the sequence,it con firmed that HCV containing in the p ositive blood of the patient is gen otype 3 and subtype展开更多
文摘1b and 2a HCV subtypes we re mainly reported in ChinaHerein ,a 3b subtype strain of HCV was fo und with gene chip methodFirstly, the 5’ NCR(non-coding region)was s elected as target sequence,and 171 related sequences in GeneBank wer e alignedAccording to the alignme nt results,eleven DNA probes were acquired and oligonucleotide array s were constructed for distinguish ing the HCV main genotypes and sub types,which did not contain specif ic probes of genotypes 4 and 5,but genotype 1, subtypes 1a/1c and 1b, genotype 2,subtypes 2a/2c and 2b,g enotype 3,subtypes 3a and 3b and g enotype 6,subtype 6aSecondly,foll owing reverse transcription of HCV RNA,nested-PCR was applied to harv est the 260 bp target DNA fragment approximatelyAfter specific ampli fication of target DNA and array h ybridization,the solid phase alkal ine phosphatase reaction indicated visually the results that the DNA fragment contains specific sequenc e of genotype 3 and subtype 3bBy sequencing the target DNA fragment ,and analyzing the sequence,it con firmed that HCV containing in the p ositive blood of the patient is gen otype 3 and subtype
