Key result
A novel 3-plex digital PCR method successfully detected HTLV-1 proviral deletions in 11.8% of asymptomatic carriers and 33.3% of high-risk carriers.
Why the study?
Two types of defective HTLV-1 proviruses are often observed in adult T-cell leukemia/lymphoma cells, motivating the development of a method to detect proviral deletions.
A novel 3-plex digital PCR method can quantitatively detect defective HTLV-1 proviruses in asymptomatic carriers and ATL patients.
New 3-plex dPCR assay detects HTLV-1 proviral deletions; leaves open clinical utility for ATL risk stratification pending validation studies.
The human T-cell leukemia virus type 1 (HTLV-1), a retrovirus, integrates into host DNA and causes adult T-cell leukemia/lymphoma (ATL) in some individuals. Two types of defective proviruses, Type 1 and Type 2, are often observed in ATL cells. Here, we developed a 3-plex digital PCR (dPCR) method to detect HTLV-1 proviral deletions by comparing the ratios of copy numbers quantified using specific primer-probes for the LTR, pol, and pX regions. We analyzed HTLV-1-positive asymptomatic carriers (ACs) and AC samples at high risk for developing ATL due to high proviral load (ATL high-risk (HR) ACs) using dPCR. Deletions were identified in 11.8 % (4/34, all Type 1) of ACs and 33.3 % (7/21, Type 1:1, Type 2:6) of ATL HR ACs. dPCR analysis revealed that in three ATL samples, all exhibited Type 1 defective characteristics, and two showed extremely low ratios in the pol region. Clonality analysis of these two samples revealed high monoclonality, indicating monoclonal expansion of ATL cells with defective proviruses. These findings demonstrate that our method effectively detects defective proviruses in both ACs and ATL, providing a valuable tool for understanding the genomic characteristics of proviruses in these conditions. • Established a 3-plex digital PCR method to detect deleted HTLV-1 proviruses. • Quantitatively detected 20 % difference in the percentage of defective proviruses. • Simple and quick detection of defective proviruses in asymptomatic carriers and ATL patients.
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Tezuka et al. (2024) studied HTLV-1 infection (n=58). 3-plex digital PCR (dPCR) method was evaluated on Detection of HTLV-1 proviral deletions. A novel 3-plex digital PCR method successfully detected HTLV-1 proviral deletions in 11.8% of asymptomatic carriers and 33.3% of high-risk carriers.
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