用于异体造血干细胞捐献脐带血筛查的细小病毒B19核酸检测法的验证

Validation of a Parvovirus B19 NAT Assay for Screening of Umbilical Cord Blood for Allogenic Hematopoietic Stem Cell Donation.

作者信息

Steininger Philipp, Korn Klaus, Hackstein Holger, Strasser Erwin F

机构信息

Institute of Clinical and Molecular Virology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.

Department of Transfusion Medicine and Haemostaseology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.

出版信息

Transfus Med Hemother. 2023 Aug 30;51(1):48-51. doi: 10.1159/000532073. eCollection 2024 Feb.

DOI:10.1159/000532073

PMID:38314242

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10836944/

Abstract

INTRODUCTION

Parvovirus B19 transmitted by umbilical cord blood (UCB) products may cause severe disease in allogenic hematopoietic stem cell transplant recipients. Thus, commercially available nucleic acid test (NAT) assays for highly sensitive detection of parvovirus B19 DNA validated for the specimen cord blood plasma (CBP) are required to avoid parvovirus B19 transmission by umbilical hematopoietic stem cell preparations.

METHODS

The multiplex cobas DPX NAT assay was validated for detection of parvovirus B19 DNA in CBP derived from citrate anticoagulated UCB units which have been processed by the Rubinstein method. In total, 363 retained CBP samples pretested negative for parvovirus B19 DNA were prepared for analyzing sensitivity, specificity, and interference of that NAT assay. The 3rd WHO International Standard for parvovirus B19 DNA was used for determining the 95% limit of detection (LOD95) by probit analysis.

RESULTS

The validation of the parvovirus B19 NAT assay for CBP demonstrated high sensitivity, specificity, intra- and inter-assay precision. Dilution series and replicate analyses showed a high linearity of the assay with a coefficient of determination above 0.99 and revealed a LOD95 of 17 International Units (IU)/mL (95% confidence interval, 14-44 IU/mL) for parvovirus B19 DNA in CBP samples.

CONCLUSION

The validation of a commercially available parvovirus B19 NAT assay for the specimen CBP demonstrated a high assay performance fulfilling German guidelines and international regulations.

摘要

引言

通过脐带血（UCB）产品传播的细小病毒B19可能会在异基因造血干细胞移植受者中引发严重疾病。因此，需要经过验证可用于高灵敏度检测脐带血血浆（CBP）样本中细小病毒B19 DNA的商用核酸检测（NAT）分析方法，以避免脐带造血干细胞制剂传播细小病毒B19。

方法

对多重cobas DPX NAT分析方法进行验证，以检测来自经鲁宾斯坦方法处理的柠檬酸盐抗凝UCB单位的CBP中的细小病毒B19 DNA。总共准备了363份预先检测细小病毒B19 DNA呈阴性的留存CBP样本，用于分析该NAT分析方法的灵敏度、特异性和干扰性情况。使用世界卫生组织第3号细小病毒B19 DNA国际标准品，通过概率分析确定95%检测限（LOD95）。