Performance of the Trioplex Real-time RT-PCR Assay for Detection of Zika, Dengue, and Chikungunya Viruses

Overview

Journal Nat Commun

Specialty Biology

Date 2018 Apr 13

PMID 29643334

Citations 93

Authors

Gilberto A Santiago

Jesus Vazquez

Sean Courtney

Katia Y Matias

Lauren E Andersen

Candimar Colon

Angela E Butler

Rebecca Roulo

John Bowzard

Julie M Villanueva

Jorge L Munoz-Jordan

Affiliations

Soon will be listed here.

Abstract

The emergence and spread of Zika virus (ZIKV) presented a challenge to the diagnosis of ZIKV infections in areas with transmission of dengue (DENV) and chikungunya (CHIKV) viruses. To facilitate detection of ZIKV infections, and differentiate these infections from DENV and CHIKV, we developed the Trioplex real-time RT-PCR assay (Trioplex assay). Here, we describe the optimization of multiplex and singleplex formats of the assay for a variety of chemistries and instruments to facilitate global standardization and implementation. We evaluated the analytical performance of all Trioplex modalities for detection of these three pathogens in serum and whole blood, and for ZIKV in urine. The limit of detection for the three viruses and in different RNA-extraction modalities is near 10 genome copy equivalents per milliliter (GCE/mL). Simultaneous testing of more than one specimen type from each patient provides a 6.4% additional diagnostic sensitivity. Overall, the high sensitivity of the Trioplex assay demonstrates the utility of this assay ascertaining Zika cases.

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References

Faye O, Faye O, Dupressoir A, Weidmann M, Ndiaye M, Sall A . One-step RT-PCR for detection of Zika virus. J Clin Virol. 2008; 43(1):96-101. DOI: 10.1016/j.jcv.2008.05.005. View

Corman V, Rasche A, Baronti C, Aldabbagh S, Cadar D, Reusken C . Assay optimization for molecular detection of Zika virus. Bull World Health Organ. 2016; 94(12):880-892. PMC: 5153932. DOI: 10.2471/BLT.16.175950. View

DOrtenzio E, Matheron S, Yazdanpanah Y, de Lamballerie X, Hubert B, Piorkowski G . Evidence of Sexual Transmission of Zika Virus. N Engl J Med. 2016; 374(22):2195-8. DOI: 10.1056/NEJMc1604449. View

Lanciotti R, Kosoy O, Laven J, Velez J, Lambert A, Johnson A . Genetic and serologic properties of Zika virus associated with an epidemic, Yap State, Micronesia, 2007. Emerg Infect Dis. 2008; 14(8):1232-9. PMC: 2600394. DOI: 10.3201/eid1408.080287. View

Reynolds M, Jones A, Petersen E, Lee E, Rice M, Bingham A . Vital Signs: Update on Zika Virus-Associated Birth Defects and Evaluation of All U.S. Infants with Congenital Zika Virus Exposure - U.S. Zika Pregnancy Registry, 2016. MMWR Morb Mortal Wkly Rep. 2017; 66(13):366-373. PMC: 5657905. DOI: 10.15585/mmwr.mm6613e1. View

Chotiwan N, Brewster C, Magalhaes T, Weger-Lucarelli J, Duggal N, Ruckert C . Rapid and specific detection of Asian- and African-lineage Zika viruses. Sci Transl Med. 2017; 9(388). PMC: 5654541. DOI: 10.1126/scitranslmed.aag0538. View

Johnson B, Russell B, Lanciotti R . Serotype-specific detection of dengue viruses in a fourplex real-time reverse transcriptase PCR assay. J Clin Microbiol. 2005; 43(10):4977-83. PMC: 1248506. DOI: 10.1128/JCM.43.10.4977-4983.2005. View

Musso D, Roche C, Robin E, Nhan T, Teissier A, Cao-Lormeau V . Potential sexual transmission of Zika virus. Emerg Infect Dis. 2015; 21(2):359-61. PMC: 4313657. DOI: 10.3201/eid2102.141363. View

Rasmussen S, Jamieson D, Honein M, Petersen L . Zika Virus and Birth Defects--Reviewing the Evidence for Causality. N Engl J Med. 2016; 374(20):1981-7. DOI: 10.1056/NEJMsr1604338. View

10.

LHuillier A, Lombos E, Tang E, Perusini S, Eshaghi A, Nagra S . Evaluation of Altona Diagnostics RealStar Zika Virus Reverse Transcription-PCR Test Kit for Zika Virus PCR Testing. J Clin Microbiol. 2017; 55(5):1576-1584. PMC: 5405276. DOI: 10.1128/JCM.02153-16. View

11.

Gyurech D, Schilling J, Schmidt-Chanasit J, Cassinotti P, Kaeppeli F, Dobec M . False positive dengue NS1 antigen test in a traveller with an acute Zika virus infection imported into Switzerland. Swiss Med Wkly. 2016; 146:w14296. DOI: 10.4414/smw.2016.14296. View

12.

Tan S, Sahoo M, Milligan S, Taylor N, Pinsky B . Stability of Zika virus in urine: Specimen processing considerations and implications for the detection of RNA targets in urine. J Virol Methods. 2017; 248:66-70. DOI: 10.1016/j.jviromet.2017.04.018. View

13.

Duffy M, Chen T, Hancock W, Powers A, Kool J, Lanciotti R . Zika virus outbreak on Yap Island, Federated States of Micronesia. N Engl J Med. 2009; 360(24):2536-43. DOI: 10.1056/NEJMoa0805715. View

14.

Bingham A, Cone M, Mock V, Heberlein-Larson L, Stanek D, Blackmore C . Comparison of Test Results for Zika Virus RNA in Urine, Serum, and Saliva Specimens from Persons with Travel-Associated Zika Virus Disease - Florida, 2016. MMWR Morb Mortal Wkly Rep. 2016; 65(18):475-8. DOI: 10.15585/mmwr.mm6518e2. View

15.

Dirlikov E, Major C, Mayshack M, Medina N, Matos D, Ryff K . Guillain-Barré Syndrome During Ongoing Zika Virus Transmission - Puerto Rico, January 1-July 31, 2016. MMWR Morb Mortal Wkly Rep. 2016; 65(34):910-4. DOI: 10.15585/mmwr.mm6534e1. View

16.

Musso D, Rouault E, Teissier A, Lanteri M, Zisou K, Broult J . Molecular detection of Zika virus in blood and RNA load determination during the French Polynesian outbreak. J Med Virol. 2016; 89(9):1505-1510. PMC: 5575554. DOI: 10.1002/jmv.24735. View

17.

Medina F, Medina J, Colon C, Vergne E, Santiago G, Munoz-Jordan J . Dengue virus: isolation, propagation, quantification, and storage. Curr Protoc Microbiol. 2012; Chapter 15:Unit 15D.2.. DOI: 10.1002/9780471729259.mc15d02s27. View

18.

Lanciotti R, Kosoy O, Laven J, Panella A, Velez J, Lambert A . Chikungunya virus in US travelers returning from India, 2006. Emerg Infect Dis. 2007; 13(5):764-7. PMC: 2738459. DOI: 10.3201/eid1305.070015. View

19.

Rabe I, Staples J, Villanueva J, Hummel K, Johnson J, Rose L . Interim Guidance for Interpretation of Zika Virus Antibody Test Results. MMWR Morb Mortal Wkly Rep. 2016; 65(21):543-6. DOI: 10.15585/mmwr.mm6521e1. View

20.

Stone M, Lanteri M, Bakkour S, Deng X, Galel S, Linnen J . Relative analytical sensitivity of donor nucleic acid amplification technology screening and diagnostic real-time polymerase chain reaction assays for detection of Zika virus RNA. Transfusion. 2017; 57(3pt2):734-747. DOI: 10.1111/trf.14031. View