Toshev Andon, Ivanova Stefka, Kovaleva Valentina, Andonova Lyubena, Mihneva Zafira
Department of Biology, Medical University of Sofia , Sofia , Bulgaria.
NCIPD, National Reference Laboratory of 'Measles, Mumps, Rubella' , Sofia , Bulgaria.
Biotechnol Biotechnol Equip. 2014 Nov 2;28(6):1103-1107. doi: 10.1080/13102818.2014.967746. Epub 2014 Oct 29.
The present study aimed to determine the involvement of the (HPVB19) as an etiological agent in individuals with fever-rash infections but not infected with rubella during the rubella outbreak (2005) in Bulgaria. A total of 194 serum samples with negative results for measles and rubella-specific IgM antibodies were tested in the National Reference Laboratory. The individuals aged 5-52 years (mean age 17.2 ± 10.15) were divided into four age groups: 5-14; 15-24; 25-34; and 35+ years old. Serological (indirect enzyme immunoassay - EIA) and molecular (extraction and detection of HPVB19-DNA) methods were used. A genotyping assay of the NS1-PCR product was proceeded with the MfeI restriction enzyme. Out of the total number of samples, 95 samples (48.97%) tested positive for HPVB19-IgM and 109 (56.18%) for HPVB19-DNA. The results from the genotyping assay revealed that genotype 1 (prototype B19) was dominant in 106 from 109 samples (97.25%), while genotype 3 (prototype V9) was detected in only 3 (2.75%). Subjects whose sera tested positive for IgM and had a positive PCR result formed a group that was most frequently linked (in 40% of cases) to acute infection. The highest prevalence was established in the group of the school-age children (5-14 years). The combined application of serological and molecular methods confirms the etiological role of HPVB19, and including virus genotyping, confirms the involvement of HPVB19 in the etiological palette of febrile rash diseases and provides a correct differential diagnostic approach.
本研究旨在确定在保加利亚2005年风疹疫情期间，人细小病毒B19（HPVB19）作为病因在发热皮疹感染但未感染风疹的个体中的作用。在国家参考实验室对194份麻疹和风疹特异性IgM抗体检测结果为阴性的血清样本进行了检测。年龄在5 - 52岁（平均年龄17.2 ± 10.15岁）的个体被分为四个年龄组：5 - 14岁；15 - 24岁；25 - 34岁；以及35岁及以上。采用了血清学方法（间接酶免疫测定 - EIA）和分子方法（HPVB19 - DNA的提取和检测）。使用MfeI限制酶对NS1 - PCR产物进行基因分型分析。在所有样本中，95份样本（48.97%）HPVB19 - IgM检测呈阳性，109份样本（56.18%）HPVB19 - DNA检测呈阳性。基因分型分析结果显示，1型（原型B19）在109份样本中的106份（97.25%）中占主导地位，而仅在3份样本（2.75%）中检测到3型（原型V9）。血清IgM检测呈阳性且PCR结果为阳性的受试者形成了一个与急性感染最常相关的组（在40%的病例中）。在学龄儿童组（5 - 14岁）中患病率最高。血清学和分子方法的联合应用证实了HPVB19的病因学作用，包括病毒基因分型，证实了HPVB19参与发热皮疹疾病的病因谱，并提供了正确的鉴别诊断方法。
