Of the 537 international migrants who consented to participate, a total of 528 (98.3%) serum samples were tested for anti-HTLV-1/2 (Murex HTLV-I + II, DiaSorin, Dartford, UK) using an enzyme-linked immunosorbent assay (ELISA). Reactive samples were confirmed using a line immunoassay (INNO-LIA HTLV I/II, Fujirebio, Europe N.V., Belgium). Samples that tested positive by LIA were considered positive for HTLV-1 or 2 infections.
Inno lia htlv 1 2
Manufactured by Fujirebio
The INNO-LIA HTLV I/II is a laboratory equipment product manufactured by Fujirebio. It is an immunoblot assay used for the detection of antibodies to human T-cell lymphotropic virus type I (HTLV-I) and type II (HTLV-II) in human serum or plasma specimens.
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4 protocols using inno lia htlv 1 2
1
Serological Screening for HTLV-1/2 Infection
2
Hepatitis C Virus and Retroviral Co-Infections
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After performing HCV load measurement using a Real-Time HCV assay (Abbott Molecular Inc., IL, USA), with a lower limit of detection of 12 IU/mL, samples were tested for HIV-1, HTLV-1 or HTLV-2 co-infection, as previously described.17
Briefly, anti-HTLV-1/-2 antibodies were detected by an enzyme immunoassay (EIA, HTLV-I/II, Gold ELISA, REM Ind. Com. Ltda, São Paulo, SP, Brazil) and confirmed by line immunoassay (INNO-LIA HTLV-I/II, Fujirebio, Europe N.V., Belgium). HIV-1 infection was evaluated by using an immunochromatographic assay (Rapid Check HIV 1 + 2, Universidade Federal do Espírito Santo, ES, Brazil) and an enzyme immunoassay (GS HIV-1/HIV-2 Plus O EIA, Bio-Rad Laboratories, USA). All assays were conducted according to the respective manufacturers’ instructions. A prevalence of 7.9% of HIV-1 and 5.3% of HTLV-1/-2 were detected, revealing HTLV-1/-2 infection in patients with hepatitis C, regardless of their HIV-1 status.17
Briefly, anti-HTLV-1/-2 antibodies were detected by an enzyme immunoassay (EIA, HTLV-I/II, Gold ELISA, REM Ind. Com. Ltda, São Paulo, SP, Brazil) and confirmed by line immunoassay (INNO-LIA HTLV-I/II, Fujirebio, Europe N.V., Belgium). HIV-1 infection was evaluated by using an immunochromatographic assay (Rapid Check HIV 1 + 2, Universidade Federal do Espírito Santo, ES, Brazil) and an enzyme immunoassay (GS HIV-1/HIV-2 Plus O EIA, Bio-Rad Laboratories, USA). All assays were conducted according to the respective manufacturers’ instructions. A prevalence of 7.9% of HIV-1 and 5.3% of HTLV-1/-2 were detected, revealing HTLV-1/-2 infection in patients with hepatitis C, regardless of their HIV-1 status.17
3
Serological Detection of HTLV-1 Antibodies
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Anti-HTLV-1 antibodies were examined by line immunoassay (INNO-LIA HTLV I/II; Fujirebio) and by particle agglutination assay (Serodia HTLV-I; Fujirebio). Plasma samples were heat inactivated at 56°C for 30 min. In INNO-LIA, 10 μL of heat-inactivated plasma samples were dispensed to 1 mL of kit-provided diluent and incubated with HTLV-1 antigen-coated strip overnight, followed by incubation with kit-provided conjugate solution and substrate solution. Then, the reaction was stopped, and the scores for relative levels of antibodies specific for individual antigens (Gag [p19 and p24] and Env [gp46 and gp21]) were determined based on the band (line) intensity according to the manufacturer's instructions. In Serodia, heat-inactivated plasma samples were serially diluted with kit-provided diluent and mixed with sensitized particles. Two hours later, the patterns of particles were evaluated. Endpoint titers were determined according to the manufacturer’s instructions. The lower limit of detection is a 1:16 dilution.
4
HTLV-1/2 Prevalence Screening in Blood Donations
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From November 2018 to March 2019, blood donations collected from Hebei, Changsha and Shenzhen blood centers were screened for HTLV-1/2 antibody using Lumipulse G HTLV-I/II Immunoreaction Cartridges set (Fujirebio, Europe, N.V, Belgium) (CLIA) on LUMIPULSE L2400 platform (Fujirebio, Europe, N.V, Belgium).At the same time, all the donations in Changsha and Hebei blood centers underwent ELISA test using Diagnostic Kit for Antibody to HTLV-1/2 (Beijing Wantai Biological Pharmacy, Beijing, China), and blood plasma samples in Shenzhen blood center were screened by Murex HTLV I + II (Diasorin, Saluggia, Italy) in respective blood center. The sample with initial reactive results were retested twice by ELISA or CLIA using the same reagent, one of the two outputs was reactive and then identified as HTLV-1/2 reactive, followed with confirmatory test using INNO-LIA HTLV I/II (Fujirebio, Europe, N.V, Belgium), the final results were analyzed by Auto-LIA 48 automatic system (Fujirebio, Europe, N.V, Belgium). The study route and Criteria for interpreting the results obtained with the INNO-LIA strips were described in Fig. 1.
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