Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation

Ueda, Miriam Yurika Hiramoto; Alvarenga, Paulo G; Real, Juliana Monte; Moreira, Eloisa De Sá; Watanabe, Aripuanã Sakurada Aranha; Passos-Castilho, Ana Maria; Vescovi, Matheus; Novis, Yana; Rocha, Vanderson; Seber, Adriana; Oliveira, José Salvador Rodrigues de; Rodrigues, Celso Arrais; Granato, Celso

doi:10.1590/0074-02760150004

Cited by 3 publications

(2 citation statements)

References 57 publications

(68 reference statements)

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“…The thermocycling conditions were as follows: 30 cycles of 95˚C for 5 min, 95˚C for 30 sec, 63˚C for 30 sec and 72˚C for 30 sec. The mRNA expression levels were normalized to GADPH using the 2 -ΔΔCq quantification method as previously described (24).…”

Section: Rna Extraction and Reverse Transcription-quantitative Polymementioning

confidence: 99%

Puerarin alters the function of monocytes/macrophages and exhibits chondroprotection in mice

et al. 2019

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Recent studies have suggested that puerarin may impede osteoclastogenesis and facilitate bone regeneration, in addition to attenuating tissue inflammation. The present study investigated the therapeutic effects of puerarin on inflammatory responses and monocyte recruitment in in vitro and in vivo osteoarthritis (OA) models. Puerarin treatment increased the proliferation of OA chondrocytes, as determined by Cell Counting Kit-8 assay. In addition, the present results suggested that puerarin suppressed the interleukin-1β-induced production of inflammatory cytokines in OA chondrocytes and monocytes/macrophages, as assessed by ELISA. In a mouse model of mono-iodoacetate-induced OA, the present histological analyses suggested that administration with puerarin attenuated the inflammatory profile of OA joints and reduced cartilage destruction. Using flow cytometry, a decreased number of myeloid-derived CC chemokine receptor 2 + /lymphocyte Ag 6C + monocytes was identified in the blood of OA mice treated with puerarin compared with control OA mice. Furthermore, quantitative real-time polymerase chain reaction analysis suggested that puerarin treatment decreased CC chemokine ligand 2 expression in arthritic tissues. Collectively, the results suggested that puerarin treatment limited the recruitment of inflammatory monocytes. In summary, the present study provided pre-clinical evidence that puerarin may serve as a potential target in the treatment of OA.

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Section: Rna Extraction and Reverse Transcription-quantitative Polymementioning

confidence: 99%

Puerarin alters the function of monocytes/macrophages and exhibits chondroprotection in mice

et al. 2019

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“…Recent studies have highlighted the importance of human herpesvirus 6 (HHV-6) in driving infection. 4 , 5 Studies have shown that nearly 40% and occasionally up to 70% of HSCT recipients develop HHV-6 infection. 6 , 7 However, the time of onset and the incidence of HHV-6 after CAR T-cell infusion have not been adequately studied.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Droplet Digital PCR and Metagenomic Next-Generation Sequencing Methods for the Detection of Human Herpesvirus 6B Infection Using Cell-Free DNA from Patients Receiving CAR-T and Hematopoietic Stem Cell Transplantation

Meng¹,

Ji²,

Chen³

et al. 2022

IDR

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Purpose The aim of this study was to examine and compare the differences between droplet digital PCR (ddPCR) and metagenomic next-generation sequencing (mNGS) in the detection of human herpesvirus 6B (HHV-6B). Long-term monitoring of HHV-6B viral load in patients receiving chimeric antigen receptor-modified T-cell (CAR-T) therapy and hematopoietic stem cell transplantation (HSCT) can be used to identify immune effector cell-associated neurotoxicity syndrome (ICANS) and guide drug therapy. Methods Twenty-seven patients with suspected HHV-6B infection who had both mNGS and ddPCR test results were analyzed retrospectively, including 19 patients who received CAR T-cell therapy and 8 who received HSCT. The HHV-6B probe and primers were designed, and the performance of the ddPCR assay was evaluated. Subsequently, ddPCR was performed utilizing blood and urine. Data on clinical information and mNGS investigations were collected. Results The ddPCR test results correlated significantly with the mNGS test results ( P < 0.001, R 2 = 0.672). Of the 27 time-paired samples, ddPCR showed positive HHV-6B detection in 20 samples, while mNGS alone showed positive HHV-6B detection in 12 samples. ddPCR detected additional HHV-6B infections in 8 samples that would have been missed if only mNGS were used. In addition, the first HHV-6B infection event was detected at a median of 14 days after CAR T-cell infusion (range, 8 to 19 days). Longitudinal monitoring of HHV-6B by ddPCR was performed to assess the effectiveness of antiviral therapy. The data showed that with antiviral treatment HHV-6B viral load gradually decreased. Conclusion Our results indicated that ddPCR improved the HHV-6B positive detection ratio and was an effective adjunct to mNGS methods. Furthermore, the longitudinal detection and quantification of HHV-6B viral load in patients undergoing CAR T-cell therapy and HSCT may serve as a guide for drug treatment.

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Comparative evaluation of a laboratory developed real-time PCR assay and the RealStar ® HHV-6 PCR Kit for quantitative detection of human herpesvirus 6

Yip

Sridhar

Cheng

et al. 2017

Journal of Virological Methods

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Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation

Cited by 3 publications

References 57 publications

Puerarin alters the function of monocytes/macrophages and exhibits chondroprotection in mice

Puerarin alters the function of monocytes/macrophages and exhibits chondroprotection in mice

Comparison of Droplet Digital PCR and Metagenomic Next-Generation Sequencing Methods for the Detection of Human Herpesvirus 6B Infection Using Cell-Free DNA from Patients Receiving CAR-T and Hematopoietic Stem Cell Transplantation

Comparative evaluation of a laboratory developed real-time PCR assay and the RealStar ® HHV-6 PCR Kit for quantitative detection of human herpesvirus 6

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