北京地区婴幼儿人类杯状病毒感染状况及型别分析

Human caliciviruses of different genotypes identified from infants and young children with acute diarrhea in Beijing area

目的 研究人类杯状病毒与北京地区婴幼儿急性腹泻的病原关系。方法 用RT PCR检测了婴幼儿非细菌性腹泻患儿粪便标本中的杯状病毒核酸 ,并用地高辛标记的不同型人类杯状病毒的cDNA(NLV基因型Ⅱ型探针CR84 0和SLV探针A14 1)作为探针 ,用斑点杂交对阳性标本进行分型。结果 从 2 33份标本中检测出阳性标本 5 8份 (阳性率 2 4 9% ) :6个月以下的幼儿阳性 19份 ,占阳性标本总数的 32 8% ,7～ 12个月的婴儿阳性 15份 ,占总阳性标本总数的 2 5 9% ,1～ 2岁的幼儿阳性9份 ,占总阳性标本总数的 15 5 % ,2～ 3岁的阳性 5份 ,占总阳性标本总数的 8 6 % ,3～ 4岁的阳性 4份 ,占总阳性标本总数的 6 9% ,5岁以上的儿童的阳性 6份 ,占总阳性标本总数的 10 2 %。杂交阳性标本共 2 9份 ,占 5 0 0 % :A14 1探针杂交阳性的标本 14份 ,占 2 4 1% ,CR84 0探针杂交阳性的标本 8份 ,占 13 8% ,A14 1和CR84 0探针杂交均为阳性的标本 7份 ,占 12 1%。结论 我国婴幼儿腹泻与人类杯状病毒感染有关 ,北京地区儿童中存在多种型别杯状病毒的感染 。

Objective To study the pathogenic relation between human caliciviruses and the acute non bacterial diarrhea in infants and young children in Beijing, China, and to determine the genotypes of human caliciviruses identified from infants and young children with acute non bacterial diarrhea Methods Human caliciviruses were detected from stool specimens collected from infants and young children with acute non bacterial diarrhea in Beijing by RT PCR using the primer pair P289/290, which can detect the nucleic acids of genotypes NLVs or SLVs Dot blot hybridization was employed to determine the genotypes of RT PCR products by digoxigenin labeled probes belonging to different genotypes which had been confirmed by sequence analysis Results Out of 233 stool specimens collected from infants and young children with acute non bacterial diarrhea seen in the affiliated children′s hospital of Capital Institute of Pediatrics, 58 (24 9%) were human calicivirus positive as determined by RT PCR The youngest infant infected with human caliciviruses was 8 days old and the oldest child infected with human caliciviruses was 12 years old Among those 58 positive specimens, 19 (32 8%) were obtained from infants younger than 6 months of age, 15 (25 9%) from those aged 7 to 12 months, 9 (15 5%) from the age group of 1 to 2 years, 5 (8 6%) were from the age group of 2 to 3 years, 4 (6 9%) from the age group of 3 to 4 years, and 6 (10 2%) were from children older than 5 years of age The probe CR840 hybridized with the PCR products obtained from stool specimen CR840 and A141and the probe A141 hybridized with the PCR products obtained from stool specimen CR840 and A141, when there was no form amide in hybridization liquid The probe CR840 did not hybridize with the PCR products obtained from stool specimen A141 and the probe A141 did not hybridize with the PCR products obtained from stool specimen CR840 until there was 50% form amide in hybridization liquid Twenty nine (50 0%) PCR products out of 58 got positive signals by dot blot hybridization with 50% form amide in hybridization liquid Among them, 14 (24 1%) hybridized with probe A141 which was obtained from a specimen from Anhui and has been determined as Sapporo like human calicivirus, 8 (13 8%) were hybridized with probe CR840 which was obtained from specimens from Beijing and has been determined as Norwalk like genotype Ⅱ, and 7 (12 1%) hybridized with both A141 and CR480 The light or dark result of hybridization was not related to the yield of RT PCR product, whereas it was related to the nucleotides identity between the probe and the RT PCR product Conclusion Human caliciviruses are related to acute non bacterial diarrhea in infants and young children in Beijing Human caliciviruses with more than two genotypes are circulating in China