Standardization of RT-qPCR Technique for Arbovirus detection in paraffin-embedded tissue samples
DOI:
https://doi.org/10.18593/evid.32787Keywords:
Arbovirus, RT-qPCR, Paraffin-embedded Tissues, Epidemiological Surveillance, Differential DiagnosisAbstract
Arboviruses are transmitted by mosquitoes and ticks, causing severe diseases in humans such as dengue, Zika, Chikungunya, and yellow fever. This study aimed to standardize the RT-qPCR technique for detecting these viruses in paraffin-embedded tissue samples. A total of 45 paraffin-embedded samples were used, including arboviruses under study and negative control samples from different periods and tissues, cataloged by the Evandro Chagas Institute. Standardization of the RT-qPCR technique is essential for arbovirus surveillance in the Amazon region and Brazil. The results demonstrated efficiency in detecting and quantifying the viral genome of different arboviruses. The RT-qPCR technique proved to be an important tool for the differential diagnosis in paraffin-embedded tissue samples. This standardization is relevant for monitoring and effectively controlling arbovirus-transmitted diseases, contributing to public health and outbreak prevention. This study highlights the importance of standardizing the RT-qPCR technique for detecting arboviruses in paraffin-embedded tissue samples. The Amazon region and Brazil are prone to outbreaks of these diseases, making these advancements even more significant. Early and accurate detection of arboviruses enables the implementation of more effective control and prevention measures, as well as contributing to the understanding of the epidemiology of these diseases.
Downloads
References
Guzman MG, Harris E. Dengue. Lancet. 2015 Jan 31;385(9966):453-65.
Thammasonthijarern N, Puangmanee W, Sriburin P, Injampa S, Chatchen S, Phumirattanaprapin W, et al. Human Heavy Chain Antibody Genes Elicited in Thai Dengue Patients during DENV2 Secondary Infection. Jpn J Infect Dis. 2020 Mar 24;73(2):140-7.
Vicenti I, Dragoni F, Giannini A, Giammarino F, Spinicci M, Saladini F, et al. Development of a Cell-Based Immunodetection Assay for Simultaneous Screening of Antiviral Compounds Inhibiting Zika and Dengue Virus Replication. SLAS Discov. 2020 Jun;25(5):506-14.
Zhou Y, Mammen MP, Klungthong C, Chinnawirotpisan P, Vaughn DW, Nimmannitya S, et al. Comparative analysis reveals no consistent association between the secondary structure of the 3’-untranslated region of dengue viruses and disease syndrome. J Gen Virol. 2006 Sep;87(Pt 9):2595-603.
Sam S-S, Omar SFS, Teoh B-T, Abd-Jamil J, AbuBakar S. Review of Dengue hemorrhagic fever fatal cases seen among adults: a retrospective study. PLoS Negl Trop Dis. 2013 May 2;7(5):e2194.
Araújo PA, Freitas MO, Chiang JO, Silva FA, Chagas LL, Casseb SM, et al. Investigation about the Occurrence of Transmission Cycles of Arbovirus in the Tropical Forest, Amazon Region. Viruses. 2019 Aug 22;11(9).
Lopez-Jimena B, Bakheit M, Bekaert M, Harold G, Frischmann S, Fall C, et al. Development and Validation of Real-Time RT-LAMP Assays for the Specific Detection of Zika Virus. Methods Mol Biol. 2020;2142:147-64.
Thompson R, Martin Del Campo J, Constenla D. A review of the economic evidence of Aedes-borne arboviruses and Aedes-borne arboviral disease prevention and control strategies. Expert Rev Vaccines. 2020 Feb;19(2):143-62.
Reed LJ, Muench H. A simple method of estimating fifty per cent endpoints. Am J Epidemiol. 1938 May;27(3):493-7.
Johnson BW, Russell BJ, Lanciotti RS. Serotype-specific detection of dengue viruses in a fourplex real-time reverse transcriptase PCR assay. J Clin Microbiol. 2005 Oct;43(10):4977-83.
Gustafsson RKL, Engdahl EE, Fogdell-Hahn A. Development and validation of a Q-PCR based TCID50 method for human herpesvirus 6. Virol J. 2012 Dec 18;9:311.
Jonsson N, Gullberg M, Lindberg AM. Real-time polymerase chain reaction as a rapid and efficient alternative to estimation of picornavirus titers by tissue culture infectious dose 50% or plaque forming units. Microbiol Immunol. 2008 Dec 18;53(3):149-54.
Guerra MJ, Ferreira CSS, Beraldo KRF, Kimura LM, Takahashi JPF, Salas-Gómez D, et al. One-step Multiplex Real-time RT-PCR for Molecular Detection and Typing of Dengue Virus Infection From Paraffin-embedded Tissues During the Brazilian 2019 Outbreak. Appl Immunohistochem Mol Morphol. 2021 Feb 1;29(2):158-62.
Corchuelo S, Gómez CY, Rosales AA, Santamaria G, Rivera JA, Saad EP, et al. CISH and IHC for the Simultaneous Detection of ZIKV RNA and Antigens in Formalin-Fixed Paraffin-Embedded Cell Blocks and Tissues. Curr Protoc. 2021 Dec;1(12):e319.
Reagan-Steiner S, Simeone R, Simon E, Bhatnagar J, Oduyebo T, Free R, et al. Evaluation of Placental and Fetal Tissue Specimens for Zika Virus Infection – 50 States and District of Columbia, January-December, 2016. MMWR Morb Mortal Wkly Rep. 2017 Jun 23;66(24):636-43.
Domingo C, Patel P, Yillah J, Weidmann M, Méndez JA, Nakouné ER, et al. Advanced yellow fever virus genome detection in point-of-care facilities and reference laboratories. J Clin Microbiol. 2012 Dec;50(12):4054-60.
Silva LA, Dermody TS. Chikungunya virus: epidemiology, replication, disease mechanisms, and prospective intervention strategies. J Clin Invest. 2017 Mar 1;127(3):737-49.
Pialoux G, Gaüzère B-A, Jauréguiberry S, Strobel M. Chikungunya, an epidemic arbovirosis. Lancet Infect Dis. 2007 May;7(5):319-27.
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Evidence
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Copyright Statement
The authors retain the copyrights and grant the Journal the right of the first publication, with the work being simultaneously licensed by a Creative Commons - Attribution - Non-Commercial 4.0 International License.
