Journal
LETTERS IN APPLIED MICROBIOLOGY
Volume 75, Issue 4, Pages 804-812Publisher
WILEY
DOI: 10.1111/lam.13752
Keywords
high resolution melting; HTLV-1; HTLV-2; nested PCR; real-time PCR; retrovirus; screening; viral diagnosis
Categories
Funding
- Italian Hospital of Buenos Aires
- CONICET [PIP 0914]
- UBACyT [20020130100783BA]
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HTLV-1 and HTLV-2 are present in different high-risk populations and have different pathogenic consequences and clinical manifestations. Accurate diagnosis of HTLV-1 and 2 infection is crucial. This study proposes a simple method for the rapid identification of HTLV-1 and 2.
HTLV-1 and HTLV-2 are present in different high-risk populations, such as sexual workers and injecting drug users (IDUs). HTLV-1 is endemic in areas of Middle East, Southern Japan and Latin America, whereas HTLV-2 infection is endemic among some Native Americans and some Central African tribes. The pathogenic consequences and clinical manifestations of these two viruses differ significantly, demanding an adequate identification; therefore, proper diagnosis of HTLV-1 and 2 infection is crucial. To get a final diagnosis of HTLV-1 or 2 infection, it is recommended that positive serologic samples should be confirmed by PCR assays or western blot (WB) analysis. Thus, the aim of this study was to develop and implement a simple reaction for the rapid identification of HTLV-1 and 2. Nested real-time PCR technique followed by high resolution melting was performed based on the tax/rex sequences of HTLV-1 (M2) and HTLV-2 (MoT) cell lines perfectly discriminating between HTLV-1 from HTLV-2, by distinct melting curve profiles. The sensitivity assay of this method revealed that at least 1 viral copy of HTLV-1 or 1 center dot 5 viral copy of HTLV-2 could be amplified. Later, this method was validated using 200 blood samples from corpses. In agreement with previous epidemiological, the HTLV-1 and 2 prevalence was 1 center dot 5% (CI 95%: 0 center dot 31-4 center dot 3) and 0 center dot 5% (CI 95%: 0 center dot 013-2 center dot 75), respectively. The strategy proposed herein has some advantages over other PCR-based tests because it not only reduces considerably time and the costs of the total diagnosis but also allows detection and discrimination of HTLV-1 and 2 in the same reaction.
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