白细胞减少患者人细小病毒B19感染的临床分子流行病学研究

Clinical molecular epidemiological study of human parvovirus B19 infection in patients with leukopenia

目的 了解白细胞减少患者人细小病毒B19（HPVB19）感染的流行状况.方法 杭州师范大学附属医院2015年1月至2016年6月前来就诊的患者,将外周血白细胞计数〈3.5×10^9/L作为入组标准,同时选取健康体检者为健康对照组.ELISA检测两组患者HPVB19 IgG和IgM,荧光定量PCR检测两组HPVB19 DNA.计数资料使用χ2检验,计量资料使用t检验.结果 共有79例患者纳入研究,男32例,女47例,年龄24～62岁;对照组126名,男55名,女71名,年龄28～67岁.白细胞减少患者的HPVB19 IgG、IgM和DNA检出率分别为34.2%、5.1%和3.8%,而健康对照组分别为36.5%、0和0,两组HPVB19 IgM和DNA检出率比较差异均有统计学意义（χ2值分别为6.507和4.856, P值分别为0.011和0.028）.对3例HPVB19 DNA阳性者进行测序分析,发现其中1例VP1/VP2序列区2个核苷酸多态性位点可以造成第153位（L/H）和219位（N/Y）氨基酸突变.系统进化分析发现2例HPVB19属于1a基因型,1例属于1b基因型.结论 白细胞减少患者的HPVB19现症感染率（HPVB19 IgM、DNA阳性）明显高于健康对照组;临床上在评估白细胞减少患者输注血制品时应检测HPVB19.

Objective To explore the distribution of parvovirus B19 （HPVB19） infection in patients with leukopenia.Methods Patients who visited the Affiliated Hospital of Hangzhou Normal University from January 2015 to June 2016 were analyzed.Patients with peripheral leucocytes count less than 3.5×109/L were included in experiment group and healthy people were included in control group.HPVB19 IgG and IgM were detected by enzyme-linked immunosorbent assay, and HPVB19 DNA was detected by quantitative polymerase chain reaction.Differences in continuous data between two groups were compared with two-sample t test and those in categorical data were compared with Chi-square test.Results A total of 79 patients were included in experiment group, including 32 males and 47 females.Ages ranged from 24 to 62 years old.And 126 healthy individuals were included in control group, including 55 males and 71 females.Ages ranged from 28 to 67 years old.The positive rates of HPVB19 IgG, IgM and DNA in experiment group were 34.2%, 5.1% and 3.8%, respectively, while those in control group were 36.5%, 0 and 0, respectively.The detection rates of HPVB19 IgM and DNA between two groups were significantly different （χ2=6.507, P=0.011 and χ2=4.856, P=0.028, respectively）.Sequence analysis for 3 of the HPVB19 DNA positive samples showed that there were two single nucleotide polymorphisms in VP1/VP2 sequence from one patient, which contributed to the 153rd （L/H） and 219th （N/Y） amino acids mutations, respectively.Phylogenetic analysis found that two strains belong to genotype 1a and one strain belongs to genotype 1b.Conclusions Detection rate of parvovirus HPVB19 infection （positive rates of HPVB19 IgM and DNA） in leukopenic patients is significantly higher than healthy controls.HPVB19 should be detected before considering transfusion in leukopenic patients in clinical practice.