Development of RT-RPA-based point-of-care tests for epidemic arthritogenic alphaviruses DOI Open Access

Sainetra Sridhar,

Prince Baffour Tonto,

Lily Lumkong

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 14, 2024

Chikungunya (CHIKV), o'nyong-nyong (ONNV), and Mayaro (MAYV) viruses are transmitted by mosquitoes known to cause a debilitating arthritogenic syndrome. These alphaviruses have emerged re-emerged, leading outbreaks in tropical subtropical regions of Asia, South America, Africa. Despite their prevalence, there persists critical gap the availability sensitive virus-specific point-of-care (POC) diagnostics. Traditional immunoglobulin-based tests such as enzyme-linked immunosorbent assay (ELISAs) often yield cross-reactive results due close genetic relationship between these viruses. Molecular diagnostics quantitative polymerase chain reaction (qPCR) offer high sensitivity but limited need for specialized laboratory equipment. Recombinase amplification (RPA), an isothermal method, is promising alternative qPCR, providing rapid with minimal equipment requirements. Here, we report development validation three RPA-based POC CHIKV, ONNV, MAYV. demonstrated both speed sensitivity, capable detecting 10 viral copies within 20 minutes amplification, without exhibiting cross-reactivity. Furthermore, evaluated clinical potential using serum tissue samples from MAYV-infected mice, well CHIKV-infected human patients. We demonstrate that RPA amplicons derived patient can be sequenced, enabling cost-effective molecular epidemiological studies. Our findings highlight significance specific improving early detection management arboviral infections.

Language: Английский

Development of an RT-RPA assay for La Crosse virus detection provides insights into age-dependent neuroinvasion in mice DOI Creative Commons

Lily Lumkong,

Reem Alatrash,

Sainetra Sridhar

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 21, 2025

Abstract La Crosse virus (LACV) is a mosquito-borne arbovirus that causes pediatric encephalitis in North America, primarily affecting children under the age of 16 years age. Early and accurate diagnosis crucial to reducing morbidity this vulnerable population; however, existing molecular serological methods face limitations sensitivity, specificity, accessibility. Here, we present development reverse transcription recombinase polymerase amplification (RT-RPA) assay for LACV detection. Our detects within 20 minutes with limit detection 100-1000 viral copies, demonstrating high specificity without cross-reactivity against closely related or geographically relevant arboviruses. We further integrate RT-RPA into lateral flow format, potentially enabling simple inexpensive point-of-care diagnosis. To complement our development, investigated pathogenesis mouse model recapitulates age-dependent susceptibility observed human populations. Using assay, reveal invades brains weanling mice as early 4 days post-infection (dpi) adult by 5 dpi. Surviving had no detectable their These results underscore utility RT-RPA-based platform offer novel insights temporal dynamics neuroinvasion clearance.

Language: Английский

Citations

0
Development of an RT-RPA assay for La Crosse virus detection provides insights into age-dependent neuroinvasion in mice DOI Creative Commons

Lily Lumkong,

Reem Alatrash,

Sainetra Sridhar

et al.

Virology Journal, Journal Year: 2025, Volume and Issue: 22(1)

Published: April 9, 2025

Language: Английский

Citations

0
Multiplex reverse transcription recombinase polymerase amplification combined with lateral flow biosensor for simultaneous detection of three viral pathogens in cattle DOI

Yating Song,

Biao Ma, Jiali Li

et al.

Talanta, Journal Year: 2024, Volume and Issue: 281, P. 126775 - 126775

Published: Aug. 30, 2024

Language: Английский

Citations

3
RT-RPA as a dual tool for detection and phylogenetic analysis of epidemic arthritogenic alphaviruses DOI Creative Commons

Sainetra Sridhar,

Prince Baffour Tonto,

Lily Lumkong

et al.

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: Dec. 3, 2024

Chikungunya (CHIKV), o'nyong-nyong (ONNV), and Mayaro (MAYV) viruses are transmitted by mosquitoes known to cause a debilitating arthritogenic syndrome. These alphaviruses have emerged re-emerged, leading outbreaks in tropical subtropical regions of Asia, South America, Africa. Despite their prevalence, there persists critical gap the availability sensitive virus-specific point-of-care (POC) diagnostics. Traditional immunoglobulin-based tests such as enzyme-linked immunosorbent assay (ELISA) often yield cross-reactive results due close genetic relationship between these viruses. Molecular diagnostics quantitative polymerase chain reaction (qPCR) offer high sensitivity but limited need for specialized laboratory equipment. Recombinase amplification (RPA), an isothermal method, is promising alternative qPCR, providing rapid with minimal equipment requirements. Here, we report development validation three RT-RPA-based CHIKV, ONNV, MAYV. demonstrated both speed sensitivity, capable detecting 10–100 viral copies within 20 min amplification, without exhibiting cross-reactivity. Furthermore, evaluated clinical potential using serum tissue samples from MAYV-infected mice, well CHIKV-infected human patients. We demonstrate that RPA amplicons derived patient can be sequenced, enabling cost-effective molecular epidemiological studies. Our findings highlight significance specific improving early detection management arboviral infections, particularly resource-limited settings.

Language: Английский

Citations

3
New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis DOI Creative Commons
Sabrina Mariotti, Maria Vincenza Chiantore,

Raffaela Teloni

et al.

Biomedicines, Journal Year: 2023, Volume and Issue: 11(2), P. 610 - 610

Published: Feb. 18, 2023

The emergence of the new pathogen SARS-CoV-2 determined a rapid need for monoclonal antibodies (mAbs) to detect virus in biological fluids as tool identify infected individuals be treated or quarantined. majority commercially available antigenic tests rely on detection N antigen biologic fluid using anti-N antibodies, and their capacity specifically subjects by is questionable due several structural analogies among proteins different coronaviruses. In order produce specific BALB/c mice were immunized three times at 20-day intervals with recombinant spike (S) protein. procedure used was highly efficient, 40 mAbs isolated, purified characterized, 13 ultimately being selected specificity lack cross reactivity other human epitopes recognized identified through peptide library and/or fragments S particular, linear along S1, excluding receptor binding domain, S2 subunits protein its major variants concern. We combinations anti-S suitable use ELISA diagnostic tests, highest sensitivity coming from proof-of-concept antigens, individuals, that represent important additional tools diagnosis COVID-19.

Language: Английский

Citations

6
Nasal swab is a good alternative sample for detecting SARS-CoV-2 with rapid antigen test: A meta-analysis DOI Creative Commons

Jia‐Wen Xie,

Ya‐Wen Zheng,

Mao Wang

et al.

Travel Medicine and Infectious Disease, Journal Year: 2023, Volume and Issue: 52, P. 102548 - 102548

Published: Feb. 9, 2023

We aim to determine if nasal samples have equivalent detection sensitivity nasopharyngeal swabs for RAT and evaluate the diagnostic accuracy of with RAT.

Language: Английский

Citations

5
Comparison of diagnostic accuracy of rapid antigen tests for COVID19 compared to the viral genetic test in adults: a systematic review and meta-analysis DOI Creative Commons
Ellyn Hirabayashi, Guadalupe Mercado,

Brandi Hull

et al.

JBI Evidence Synthesis, Journal Year: 2024, Volume and Issue: 22(10), P. 1939 - 2002

Published: Aug. 26, 2024

The objective of this review was to determine the diagnostic accuracy currently available and upcoming point-of-care rapid antigen tests (RATs) used in primary care settings relative viral genetic real-time reverse transcriptase polymerase chain reaction (RT-PCR) test as a reference for diagnosing COVID-19/SARS-CoV-2 adults.

Language: Английский

Citations

1
Performance of self-performed SARS-CoV-2 rapid antigen test: a systematic review and meta-analysis DOI Creative Commons
Peiling Cai, Junren Wang, Peng Ye

et al.

Frontiers in Public Health, Journal Year: 2024, Volume and Issue: 12

Published: Oct. 18, 2024

The aim of this study was to investigate the accuracy self-tested SARS-CoV-2 rapid antigen tests.

Language: Английский

Citations

1
Update of European Society of Clinical Microbiology and Infectious Diseases coronavirus disease 2019 guidelines: diagnostic testing for severe acute respiratory syndrome coronavirus 2 DOI Creative Commons
Paraskevi C. Fragkou, Giulia De Angelis, Giulia Menchinelli

et al.

Clinical Microbiology and Infection, Journal Year: 2023, Volume and Issue: 29(7), P. 876 - 886

Published: April 23, 2023

Language: Английский

Citations

3
Optimal targeted mass screening in non‐uniform populations with multiple tests and schemes DOI
Jiayi Lin, Hrayer Aprahamian, George Golovko

et al.

Naval Research Logistics (NRL), Journal Year: 2023, Volume and Issue: 71(1), P. 87 - 108

Published: Aug. 3, 2023

Abstract We study the problem of designing optimal targeted mass screening non‐uniform populations. Mass is an essential tool that widely utilized in a variety settings, for example, preventing infertility through programs sexually transmitted diseases, ensuring safe blood supply transfusion, and mitigating transmission infectious diseases. The objective to maximize overall classification accuracy under limited budget. In this paper, we address by proposing proactive optimization‐based framework factors population heterogeneity, budget, different testing schemes, availability multiple assays, imperfect assays. By analyzing resulting optimization problem, take advantage structure as multi‐dimensional fractional knapsack identify efficient globally convergent threshold‐style solution scheme fully characterizes across entire budget spectrum. Using real‐world data, conduct geographic‐based nationwide case on COVID‐19 United States. Our results reveal identified strategies substantially outperform conventional practices significantly lowering misclassifications while utilizing same amount Moreover, our provide valuable managerial insights with regard distribution geographic regions.

Language: Английский

Citations

1