Development and Validation of a Rapid Lateral Flow E1/E2-Antigen Test and ELISA in Patients Infected with Emerging Asian Strain of Chikungunya Virus in the Americas.

Ankita Reddy,Nol Salcedo,Natalia Andrea Gómez Ríos,Diana María Caicedo-Borrero,Bobby Brooke Herrera,Marcela Mercado,Helena De Puig,Irene Bosch,Michael S Diamond,Margarita Gélvez-Ramírez,Luis A Villar-Centeno,Leda Parham,Kimberly García,Lee Gehrke,Angélica María Rico Turca,Carlos F Narváez,Ivette Lorenzana,Dawlyn García,Megan Hiley

doi:10.3390/v12090971

Abstract

Since its 2013 emergence in the Americas, Chikungunya virus (CHIKV) has posed a serious threat to public health. Early and accurate diagnosis of the disease, though currently lacking in clinics, is integral to enable timely care and epidemiological response. We developed a dual detection system: a CHIKV antigen E1/E2-based enzyme-linked immunosorbent assay (ELISA) and a lateral flow test using high-affinity anti-CHIKV antibodies. The ELISA was validated with 100 PCR-tested acute Chikungunya fever samples from Honduras. The assay had an overall sensitivity and specificity of 51% and 96.67%, respectively, with accuracy reaching 95.45% sensitivity and 92.03% specificity at a cycle threshold (Ct) cutoff of 22. As the Ct value decreased from 35 to 22, the ELISA sensitivity increased. We then developed and validated two lateral flow tests using independent antibody pairs. The sensitivity and specificity reached 100% for both lateral flow tests using 39 samples from Colombia and Honduras at Ct cutoffs of 20 and 27, respectively. For both lateral flow tests, sensitivity decreased as the Ct increased after 27. Because CHIKV E1/E2 are exposed in the virion surfaces in serum during the acute infection phase, these sensitive and specific assays demonstrate opportunities for early detection of this emerging human pathogen.

Highlights

Chikungunya virus (CHIKV) is an increasingly prevalent alphavirus that is transmitted by Aedes mosquitoes [1]
Of 1056 antibodies harvested from the CHIKV-immunized mice, mAb 48 with mAb 155 (Combination A) and mAb 4 with mAb 340 (Combination B) were selected for the sandwich enzyme-linked immunosorbent assay (ELISA)
We show that of the ELISA positive clones, 16 out of 41 clones tested positive using fluorescent-activated cell sorting (FACS), corresponding to 39% positivity (Figure S2)

Summary

Introduction

Chikungunya virus (CHIKV) is an increasingly prevalent alphavirus that is transmitted by Aedes mosquitoes [1]. CHIKV has re-emerged at an unprecedented rate, spreading to over 100 countries across five continents and producing over one million infections annually [2,3,4]. As mosquito breeding grounds expand in response to climate change and globalization, CHIKV infections are expected to pose an even greater threat to public health. Chikungunya fever, which is caused by infection of CHIKV, is a debilitating disease which often includes joint pain and high fever, and a plethora of additional nonspecific symptoms including rash, abdominal pain, vomiting, diarrhea, myalgia, and headache. The acute clinical manifestations often subside after 1−3 weeks, chronic joint pain lasting months to years can significantly impair movement and undermine quality of life [5]. Severe forms of Chikungunya fever include neurological complications, myocarditis, pneumonia, lymphadenopathy, hepatitis, and pancreatitis [2]

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