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Tytuł pozycji:

Use of synthetic oligonucleotides for determination of HTLV-1 proviral load by real-time PCR: a helpful alternative approach in the clinical management.

Tytuł :
Use of synthetic oligonucleotides for determination of HTLV-1 proviral load by real-time PCR: a helpful alternative approach in the clinical management.
Autorzy :
Bandeira LM; Federal University of Mato Grosso do Sul, Campo Grande, MS, Brazil.
Puga MAM; Federal University of Mato Grosso do Sul, Campo Grande, MS, Brazil.
de Paula VS; Oswaldo Cruz Institute, Rio de Janeiro, RJ, Brazil.
Demarchi LHF; Central Public Health Laboratory, Lacen/MS, Campo Grande, MS, Brazil.
Lichs GGC; Central Public Health Laboratory, Lacen/MS, Campo Grande, MS, Brazil.
Domingos JA; Federal University of Mato Grosso do Sul, Campo Grande, MS, Brazil.
da Cunha RV; Federal University of Mato Grosso do Sul, Campo Grande, MS, Brazil.; Oswaldo Cruz Foundation, Campo Grande, MS, Brazil.
Uehara SNO; Federal University of Mato Grosso do Sul, Campo Grande, MS, Brazil.
Motta-Castro ARC; Federal University of Mato Grosso do Sul, Campo Grande, MS, Brazil.; Oswaldo Cruz Foundation, Campo Grande, MS, Brazil.
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Źródło :
Journal of applied microbiology [J Appl Microbiol] 2020 Sep; Vol. 129 (3), pp. 768-774. Date of Electronic Publication: 2020 Apr 08.
Typ publikacji :
Journal Article
Język :
Imprint Name(s) :
Original Publication: Oxford : Published for the Society for Applied Bacteriology by Blackwell Science, c1997-
MeSH Terms :
HTLV-I Infections/*diagnosis
Human T-lymphotropic virus 1/*isolation & purification
Proviruses/*isolation & purification
Real-Time Polymerase Chain Reaction/*methods
Viral Load/*methods
Adult ; DNA, Viral/genetics ; Disease Progression ; Genome, Viral/genetics ; HTLV-I Infections/blood ; HTLV-I Infections/virology ; Human T-lymphotropic virus 1/genetics ; Humans ; Leukocytes, Mononuclear/virology ; Middle Aged ; Oligonucleotides/chemical synthesis ; Oligonucleotides/genetics ; Prognosis ; Proviruses/genetics ; Real-Time Polymerase Chain Reaction/standards ; Viral Load/standards
References :
Bandeira, L.M., Uehara, S.N., Asato, M.A., Aguena, G.S., Maedo, C.M., Benites, N.H., Puga, M.A., Rezende, G.R. et al. (2015) High prevalence of HTLV-1 infection among Japanese immigrants in non-endemic area of Brazil. PLoS Negl Trop Dis 9, e0003691.
Bustin, S.A., Benes, V., Garson, J.A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T. et al. (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55, 611-622.
Conte, J., Potoczniak, M.J. and Tobe, S.S. (2018) Using synthetic oligonucleotides as standards in probe-based qPCR. Biotechniques 64, 177-179.
Dehée, A., Césaire, R., Désiré, N., Lézin, A., Bourdonné, O., Béra, O., Plumelle, Y., Smadja, D. et al. (2002) Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay. J Virol Methods 102, 37-51.
Furtado, M.D.O.S.S., Andrade, R.G., Romanelli, L.C., Ribeiro, M.A., Ribas, J.G., Torres, E.B., Barbosa-Stancioli, E.F., Proietti, A.B. et al. (2012) Monitoring the HTLV-1 proviral load in the peripheral blood of asymptomatic carriers and patients with HTLV-associated myelopathy/tropical spastic paraparesis from a Brazilian cohort: ROC curve analysis to establish the threshold for risk disease. J Med Virol 84, 664-671.
Grassi, M.F., Olavarria, V.N., Kruschewsky, R.D., Yamano, Y., Jacobson, S., Taylor, G.P., Martin, F. and Galvão-Castro, B. (2013) Utility of HTLV proviral load quantification in diagnosis of HTLV-1-associated myelopathy requires international standardization. J Clin Virol 58, 584-586.
Lima, L.R.P., Silva, A.P., Schmidt-Chanasit, J. and de Paula, V.S. (2017) Diagnosis of human herpes virus 1 and 2 (HHV-1 and HHV-2): use of a synthetic standard curve for absolute quantification by real time polymerase chain reaction. Mem Inst Oswaldo Cruz 112, 220-223.
Martinez-Martinez, M., Diez-Valcarce, M., Hernández, M. and Rodríguez-Lázaro, D. (2011) Design and application of nucleic acid standards for quantitative detection of enteric viruses by real-time PCR. Food Environ Virol 3, 92-98.
Martins, M.L., de Freitas Carneiro-Proietti, A.B., Nicolato, R., de Miranda, D.M. and Romanelli, L.C.F. (2018) HTLV-1 proviral load in cerebrospinal fluid may not be a good marker to differentiate asymptomatic carriers with high proviral load in blood from HAM/TSP patients. J Neurovirol 24, 432-438.
Mayne, M., Cheadle, C., Soldan, S.S., Cermelli, C., Yamano, Y., Akhyani, N., Nagel, J.E., Taub, D.D. et al. (2001) Gene expression profile of herpesvirus-infected T cells obtained using immunomicroarrays: induction of proinflammatory mechanisms. J Virol 75, 11641-11650.
Nagai, M., Usuku, K., Matsumoto, W., Kodama, D., Takenouchi, N., Moritoyo, T., Hashiguchi, S., Ichinose, M. et al. (1998) Analysis of HTLV-I proviral load in 202 HAM/TSP patients and 243 asymptomatic HTLV-I carriers: high proviral load strongly predisposes to HAM/TSP. J Neurovirol 4, 586-593.
Paiva, A.M., Assone, T., Haziot, M.E.J., Smid, J., Fonseca, L.A.M., Luiz, O.D.C., de Oliveira, A.C.P. and Casseb, J. (2018) Risk factors associated with HTLV-1 vertical transmission in Brazil: longer breastfeeding, higher maternal proviral load and previous HTLV-1-infected offspring. Sci Rep 8, 7742.
Portilho, M.M., Mendonça, A.C.D.F., Bezerra, C.S., do Espirito-Santo, M.P., de Paula, V.S., Nabuco, L.C., Villela-Nogueira, C.A., Lewis-Ximenez, L.L. et al. (2018) Usefulness of in-house real time PCR for HBV DNA quantification in serum and oral fluid samples. J Virol Methods 256, 100-106.
Rosadas, C., Cabral-Castro, M.J., Vicente, A.C., Peralta, J.M. and Puccioni-Sohler, M. (2013) Validation of a quantitative real-time PCR assay for HTLV-1 proviral load in peripheral blood mononuclear cells. J Virol Methods 193, 536-541.
Shoeibi, A., Etemadi, M.M., Moghaddam Ahmadi, A., Amini, M. and Boostani, R. (2013) “HTLV-I Infection” twenty-year research in neurology department of Mashhad University of Medical Sciences. Iran J Basic Med Sci 16, 202-207.
Tamegão-Lopes, B.P., Rezende, P.R., Maradei-Pereira, L.M.C. and de Lemos, J.A.R. (2006) HTLV-1 and HTLV-2 proviral load: a simple method using quantitative real-time PCR. Rev Soc Bras Med Trop 39, 548-552.
Thulin Hedberg, S., Eriksson, L., Demontis, M.A., Mölling, P., Sundqvist, M., Taylor, G., Malm, K. and Andersson, S. (2018) Droplet digital PCR for absolute quantification of proviral load of human T-cell lymphotropic virus (HTLV) types 1 and 2. J Virol Methods 260, 70-74.
Tourinho, R., de Almeida, C., Lemos, A., Gardinali, N.R., Vieira, Y.R., Schmidt-Chanasit, J. and de Paula, V.S. (2015) Application of synthetic standard curves for absolute quantification of hepatitis A and E by real-time PCR. J Genet Genome Res 2, 013.
Verdonck, K., González, E., Van Dooren, S., Vandamme, A.M., Vanham, G. and Gotuzzo, E. (2007) Human T-lymphotropic vírus 1: recent knowledge about an ancient infection. Lancet Infect Dis 7, 266-281.
Zihlmann, K.F., de Alvarenga, A.T. and Casseb, J. (2012) Living invisible: HTLV-1-infected persons and the lack of care in public health. PLoS Negl Trop Dis 6, e1705.
Grant Information :
Fundação Oswaldo Cruz; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Contributed Indexing :
Keywords: HTLV; HTLV-1; HTLV-1 proviral load; qPCR; quantification; synthetic oligonucleotides standard curve
Substance Nomenclature :
0 (DNA, Viral)
0 (Oligonucleotides)
Entry Date(s) :
Date Created: 20200324 Date Completed: 20201019 Latest Revision: 20201019
Update Code :
Czasopismo naukowe
Aims: To evaluate the potential use of synthetic oligonucleotides as a standard curve for proviral load (PVL) of human T-cell leukaemia virus type 1 (HTLV-1) quantification in peripheral blood mononuclear cells (PBMC) of HTLV-1-infected individuals by quantitative real-time polymerase chain reaction (qPCR) analysis.
Methods and Results: Synthetic oligonucleotides based on HTLV-1 genome were customized to use as a standard curve. Twelve anti-HTLV-1-positive samples with known HTLV-1 PVL, previously quantified by qPCR assay using TARL-2 cells as a conventional standard curve, were submitted to the new protocol. The proviral quantification levels had a high concordance with qPCR results using a conventional standard curve. The results demonstrate that the conventional standard curve can be replaced by a synthetic standard curve due to its ability to quantification based on the linearity and qPCR efficiency and similar results with a validated qPCR assay using a conventional standard curve.
Conclusions: Synthetic oligonucleotides standard curves could be a very useful tool on HTLV-1 diagnosis and absolute HTLV-1 PVL quantification.
Significance and Impact of the Study: HTLV-1 PVL determination using synthetic oligonucleotides standard curve by qPCR could be a helpful alternative for the laboratories that monitor infected patients as an important prognostic factor in HTLV-1-associated diseases progression. Also, it can decrease costs and overcome the biological limitations of the plasmid curve.
(© 2020 The Society for Applied Microbiology.)

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