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Feasibility of the Routine Clinical Use of a Multiplex Virus Polymerase Chain Reaction Assay Based on Blood Virus Detection in Hematopoietic Stem Cell-Transplanted Patients

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International Journal of Clinical Medicine 2022年第2期13卷 67-81页

作者： Hiroko Tsunemine Miho Sasaki Yuriko Zushi Toshiharu Saitoh Norio Shimizu Yasuhiro Tomaru Yumi Aoyama Ryusuke Yamamoto Tomomi Sakai nobuyoshi arima Taiichi Kodaka Takayuki Takahashi Departments of Hematology Shinko Hospital Kobe Japan Cell Therapy Shinko Hospital Kobe Japan Center for Stem Cell and Regenerative Medicine Tokyo Medical and Dental University Tokyo Japan Akasaka Hematology & Oncology Clinic Kobe Japan

Background: Multiplex virus assays are useful in immunocompromised hosts but still challenging in routine clinical settings in terms of their sensitivity, specificity, reproducibility, and time and cost performances. In recent years, we developed a qualitative multiplex virus PCR assay capable of the simultaneous detection of 13 virus species within 3 h. However, because of the multiple and concomitant nature of this virus assay, it should be validated for qualitative reliability. Materials and Methods: As a preclinical examination, this multiplex PCR was able to detect 1.25 × 103 copies/mL of 13 synthesized virus genomes and preserved same virus DNAs by the serial dilution method. Blood samples from 40 patients who underwent hematopoietic stem cell transplantation were then examined by multiplex PCR for 13 virus species, followed by quantitative real-time PCR for all 13 virus species as reference PCR when these patients developed symptoms suggestive of viral infection. Results: In 421 cumulative qualitative-quantitative tests, the multiplex PCR certainly detected 1.0 × 103 copies/mL of 5 viruses (CMV, JCV, BKV, HHV-6, ADV) that were frequently detected and thus reasonably analyzed. The positive and negative predictive values of multiplex PCR were 84.2% - 93.3% and 90.7% - 99.0%, respectively, and sensitivity and specificity were 59.0% - 83.3% and 97.2% - 99.2%, respectively, for these 5 viruses. Conclusion: From these performances, the multiplex PCR assay may be acceptable in a routine clinical laboratory setting.

关键词： Multiplex Virus PCR Assay Routine Laboratory Use Positive Predictive Value Negative Predictive Value Sensitivity Specificity

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