Establishment of an Alphavirus-Specific Neutralization Assay to Distinguish Infections with Different Members of the Semliki Forest Complex

Abstract

Background: Alphaviruses are transmitted by arthropod vectors and can be found worldwide. Alphaviruses of the Semliki Forest complex such as chikungunya virus (CHIKV), Mayaro virus (MAYV) or Ross River virus (RRV) cause acute febrile illness and long-lasting arthralgia in humans, which cannot be clinically discriminated from a dengue virus or Zika virus infection. Alphaviruses utilize a diverse array of mosquito vectors for transmission and spread. For instance, adaptation of CHIKV to transmission by Aedes albopictus has increased its spread and resulted in large outbreaks in the Indian Ocean islands. For many alphaviruses commercial diagnostic tests are not available or show cross-reactivity among alphaviruses. Climate change and globalization will increase the spread of alphaviruses and monitoring of infections is necessary and requires virus-specific methods. Method: We established an alphavirus neutralization assay in a 384-well format by using pseudotyped lentiviral vectors. Results: MAYV-specific reactivity could be discriminated from CHIKV reactivity. Human plasma from blood donors infected with RRV could be clearly identified and did not cross-react with other alphaviruses. Conclusion: This safe and easy to use multiplex assay allows the discrimination of alphavirus-specific reactivity within a single assay and has potential for epidemiological surveillance. It might also be useful for the development of a pan-alphavirus vaccine.

Arthritogenic Alphaviruses: A Worldwide Emerging Threat?

“…Regardless, cross-reactivity remains a main limitation in biological diagnostics, especially for immunological tests [274,275,276]. Hence, the closer alphaviruses are phylogenetically, the more cross-reactivity exists [194]. In response, considerable effort is made on developing technologies to avoid cross-reactivity both in direct [271,277] and indirect diagnosis [194,278].…”

“…Hence, the closer alphaviruses are phylogenetically, the more cross-reactivity exists [194]. In response, considerable effort is made on developing technologies to avoid cross-reactivity both in direct [271,277] and indirect diagnosis [194,278].…”

“…The main pitfall of serological methods is the cross-reactivity among alphaviruses that complicates identifying which of the arthritogenic alphaviruses is involved, especially in regions where they co-circulate. For this reason, the plaque-reduction neutralizing test (PRNT) remains a gold standard for diagnostics [193,194].…”

More Than Just Gene Therapy Vectors: Lentiviral Vector Pseudotypes for Serological Investigation

“…There are many other examples in the emerging virus field where a pseudotyped LV has been used to investigate NAbs induced by candidate vaccines. Chikungunya virus-pseudotyped LVs have been used to determine the level of NAbs raised in non-human primates by a virus-like particle vaccine candidate and showed good correlation between the pseudotyped LV neutralisation method and the traditional PRNT with the live virus [ 88 ]; similar results were obtained using a Lassa fever virus pseudotyped LV for the evaluation of a candidate DNA vaccine in a non-human primate model [ 21 ]. This has further been demonstrated with Ebola virus pseudotyped LVs as part of a phase I clinical trial of a viral vectored vaccine regime [ 49 ].…”

Inactivation and Removal of Chikungunya Virus and Mayaro Virus from Plasma-derived Medicinal Products

“…The close phylogenetic relationship amongst members of the Semliki Forest complex complicates specific antibody detection. As a consequence, few alphavirus-specific diagnostic kits are commercially available, some of them showing inevitable cross-reactivity [10], in particular between MAYV, CHIKV and o’nyong-nyong virus [11,12]. Common serological assays such as plaque reduction neutralization tests (PRNT 50 ) or reduction of cytopathic effects in a 96-well plate format (tissue culture infectious dose, TCID 50 ), which both detect antibodies specifically neutralizing infectious virus particles, require pre-treatment of the plasma/serum samples at 56 °C for 30 min to inactivate complement and adventitious viruses which could interfere with the serological test system.…”