Development of reference material with assigned value for human T‐cell leukemia virus type 1 quantitative PCR in Japan
M Kuramitsu, K Okuma, M Nakashima… - Microbiology and …, 2018 - Wiley Online Library
M Kuramitsu, K Okuma, M Nakashima, T Sato, D Sasaki, H Hasegawa, K Umeki, R Kubota…
Microbiology and immunology, 2018•Wiley Online LibraryQuantitative PCR (qPCR) of human T‐cell leukemia virus type 1 (HTLV‐1) provirus is used
for HTLV‐1 testing and for assessment of risk of HTLV‐1‐related diseases. In this study, a
reference material was developed for standardizing HTLV‐1 qPCR. Freeze‐dried TL‐Om1
cells diluted with Jurkat cells were prepared and an assigned value for proviral load (PVL) of
2.71 copies/100 cells was determined by digital PCR. Nine Japanese laboratories using
their own methods evaluated the PVLs of this reference material as 1.08–3.49 copies/100 …
for HTLV‐1 testing and for assessment of risk of HTLV‐1‐related diseases. In this study, a
reference material was developed for standardizing HTLV‐1 qPCR. Freeze‐dried TL‐Om1
cells diluted with Jurkat cells were prepared and an assigned value for proviral load (PVL) of
2.71 copies/100 cells was determined by digital PCR. Nine Japanese laboratories using
their own methods evaluated the PVLs of this reference material as 1.08–3.49 copies/100 …
Abstract
Quantitative PCR (qPCR) of human T‐cell leukemia virus type 1 (HTLV‐1) provirus is used for HTLV‐1 testing and for assessment of risk of HTLV‐1‐related diseases. In this study, a reference material was developed for standardizing HTLV‐1 qPCR. Freeze‐dried TL‐Om1 cells diluted with Jurkat cells were prepared and an assigned value for proviral load (PVL) of 2.71 copies/100 cells was determined by digital PCR. Nine Japanese laboratories using their own methods evaluated the PVLs of this reference material as 1.08–3.49 copies/100 cells. The maximum difference between laboratories was 3.2‐fold. Correcting measured PVLs by using a formula incorporating the assigned value of this reference material should minimize such discrepancies.
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