Laboratory evaluation of RealStar Yellow Fever Virus RT-PCR kit 1.0 for potential use in the global yellow fever laboratory network
Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always...
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| Vydáno v: | PLoS neglected tropical diseases Ročník 16; číslo 9; s. e0010770 |
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| Médium: | Journal Article |
| Jazyk: | angličtina |
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Public Library of Science
01.09.2022
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| ISSN: | 1935-2735, 1935-2727, 1935-2735 |
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| Abstract | Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0.
Specific objectives were to assess the assay's ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly.
The RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer's claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives. |
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| AbstractList | Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0.BACKGROUNDEarly detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0.Specific objectives were to assess the assay's ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly.METHODOLOGY AND PRINCIPAL FINDINGSSpecific objectives were to assess the assay's ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly.The RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer's claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives.CONCLUSIONS AND SIGNIFICANCEThe RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer's claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives. Background Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0. Methodology and principal findings Specific objectives were to assess the assay’s ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly. Conclusions and significance The RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer’s claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives. Background Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0. Methodology and principal findings Specific objectives were to assess the assay’s ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly. Conclusions and significance The RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer’s claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives. BackgroundEarly detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0.Methodology and principal findingsSpecific objectives were to assess the assay's ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly.Conclusions and significanceThe RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer's claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives. Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0. Specific objectives were to assess the assay's ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly. The RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer's claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives. Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly rely on serological assays that require confirmation at Regional Reference Laboratories, thus delaying results, which themselves are not always definitive often due to antibody cross-reactivity. A positive molecular test result is confirmatory for YF; therefore, a standardized YF molecular assay would facilitate immediate confirmation at National Laboratories. The WHO-coordinated global Eliminate Yellow Fever Epidemics Laboratory Technical Working Group sought to independently evaluate the quality and performance of commercial YF molecular assays relevant to use in countries with endemic YF, in the absence of stringent premarket assessments. This report details a limited laboratory WHO-coordinated evaluation of the altona Diagnostics RealStar Yellow Fever Virus RT-PCR kit 1.0. Specific objectives were to assess the assay's ability to detect YF virus strains in human serum from YF-endemic regions, determine the potential for interference and cross-reactions, verify the performance claims as stated by the manufacturer, and assess usability. RNA extracted from normal human serum spiked with YF virus showed the assay to be precise with minimal lot-to-lot variation. The 95% limit of detection calculated was approximately 1,245 RNA copies/ml [95% confidence interval 497 to 1,640 copies/ml]. Positive results were obtained with spatially and temporally diverse YF strains. The assay was specific for YF virus, was not subject to endogenous or exogenous interferents, and was clinically sensitive and specific. A review of operational characteristics revealed that a positivity cutoff was not defined in the instructions for use, but otherwise the assay was user-friendly. The RealStar Yellow Fever Virus RT-PCR kit 1.0 has performance characteristics consistent with the manufacturer's claims and is suitable for use in YF-endemic regions. Its use is expected to decrease YF outbreak detection times and be instrumental in saving lives. Molecular diagnosis of human specimens is the fastest way to confirm and subsequently mitigate yellow fever (YF) outbreaks. In Africa, a serological method requiring lengthy confirmation external to the National Laboratories is standard and results are not always definitive due to factors such as cross-reactive antibodies. Molecular testing has been uncommon in African YF laboratories, largely due to challenges with acquiring reagents and a lack of an approved assay that simplifies laboratory procedures. In our capacities as members of the global Eliminate Yellow Fever Epidemics strategy, we independently evaluated the RealStar Yellow Fever RT-PCR kit 1.0 (altona Diagnostics, Hamburg, Germany). We showed that the assay is sensitive and specific for detection of RNA from representative YF virus strains found worldwide. It is not subject to interference from common blood contaminants and has good clinical performance. In conjunction with a separately conducted review of the manufacturer’s quality systems, we concluded that the RealStar Yellow Fever Virus RT-PCR kit 1.0 is appropriate for use in YF endemic regions. Logistical and funding support to introduce the assay is being arranged, which should decrease YF outbreak detection times and ultimately save lives. |
| Audience | Academic |
| Author | Brault, Aaron C. Goodman, Christin H. Basile, Alison J. Johnson, Barbara W. Velez, Jason O. Domingo, Cristina Mossel, Eric C. Hughes, Holly R. Meurant, Robyn Niedrig, Matthias Lambert, Amy J. Mulders, Mick N. |
| AuthorAffiliation | 6 World Health Organization, Geneva, Switzerland 1 Arboviral Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, United States of America 2 WHO consultant, Berlin, Germany 4 Public Health Laboratory Support Unit, Centre for International Health Protection, Robert Koch Institute, Berlin, Germany 5 Scientific Laboratory Consulting, Laporte, Colorado, United States of America 3 ACT-IVD, Bougival, France Aix-Marseille Universite, FRANCE |
| AuthorAffiliation_xml | – name: 2 WHO consultant, Berlin, Germany – name: 5 Scientific Laboratory Consulting, Laporte, Colorado, United States of America – name: Aix-Marseille Universite, FRANCE – name: 6 World Health Organization, Geneva, Switzerland – name: 3 ACT-IVD, Bougival, France – name: 4 Public Health Laboratory Support Unit, Centre for International Health Protection, Robert Koch Institute, Berlin, Germany – name: 1 Arboviral Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, United States of America |
| Author_xml | – sequence: 1 givenname: Alison J. surname: Basile fullname: Basile, Alison J. – sequence: 2 givenname: Matthias surname: Niedrig fullname: Niedrig, Matthias – sequence: 3 givenname: Amy J. surname: Lambert fullname: Lambert, Amy J. – sequence: 4 givenname: Robyn surname: Meurant fullname: Meurant, Robyn – sequence: 5 givenname: Aaron C. surname: Brault fullname: Brault, Aaron C. – sequence: 6 givenname: Cristina surname: Domingo fullname: Domingo, Cristina – sequence: 7 givenname: Christin H. surname: Goodman fullname: Goodman, Christin H. – sequence: 8 givenname: Barbara W. surname: Johnson fullname: Johnson, Barbara W. – sequence: 9 givenname: Eric C. surname: Mossel fullname: Mossel, Eric C. – sequence: 10 givenname: Mick N. surname: Mulders fullname: Mulders, Mick N. – sequence: 11 givenname: Jason O. surname: Velez fullname: Velez, Jason O. – sequence: 12 givenname: Holly R. orcidid: 0000-0003-1380-8263 surname: Hughes fullname: Hughes, Holly R. |
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| DOI | 10.1371/journal.pntd.0010770 |
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| References_xml | – ident: pntd.0010770.ref021 – volume: 56 issue: 10 year: 2018 ident: pntd.0010770.ref012 article-title: Yellow Fever Virus: Diagnostics for a Persistent Arboviral Threat publication-title: Journal of clinical microbiology doi: 10.1128/JCM.00827-18 – ident: pntd.0010770.ref015 – volume: 15 start-page: 177 issue: 2 year: 2008 ident: pntd.0010770.ref017 article-title: Evaluation of an indirect immunofluorescence assay for detection of immunoglobulin M (IgM) and IgG antibodies against yellow fever virus publication-title: Clin Vaccine Immunol doi: 10.1128/CVI.00078-07 – ident: pntd.0010770.ref023 – ident: pntd.0010770.ref001 – volume: 4 start-page: 503 year: 1976 ident: pntd.0010770.ref018 article-title: Serum dilution neutralization test for California group virus identification and serology publication-title: Journal of clinical microbiology doi: 10.1128/jcm.4.6.503-510.1976 – volume: 38 start-page: 402 year: 2021 ident: pntd.0010770.ref013 article-title: Laboratory capacity assessments in 25 African countries at high risk of yellow fever, August-December 2018 publication-title: Pan Afr Med J doi: 10.11604/pamj.2021.38.402.28886 – volume: 50 start-page: 4054 issue: 12 year: 2012 ident: pntd.0010770.ref020 article-title: Advanced yellow fever virus genome detection in point-of-care facilities and reference laboratories publication-title: Journal of clinical microbiology doi: 10.1128/JCM.01799-12 – ident: pntd.0010770.ref024 – volume: 381 start-page: 444 issue: 5 year: 2019 ident: pntd.0010770.ref007 article-title: Immunogenicity of Fractional-Dose Vaccine during a Yellow Fever Outbreak—Final Report publication-title: N Engl J Med doi: 10.1056/NEJMoa1710430 – volume: 64 start-page: 160 year: 2015 ident: pntd.0010770.ref002 article-title: Yellow fever publication-title: J Clin Virol doi: 10.1016/j.jcv.2014.08.030 – year: 2016 ident: pntd.0010770.ref014 – volume: 3 start-page: e75 issue: 5 year: 2007 ident: pntd.0010770.ref003 article-title: Out of Africa: a molecular perspective on the introduction of yellow fever virus into the Americas publication-title: PLoS Pathog doi: 10.1371/journal.ppat.0030075 – ident: pntd.0010770.ref022 – volume: 276 start-page: 1157 issue: 14 year: 1996 ident: pntd.0010770.ref011 article-title: Yellow Fever: A Decade of Reemergence publication-title: JAMA doi: 10.1001/jama.1996.03540140045025 – volume: 394 start-page: 12 issue: 1 year: 2009 ident: pntd.0010770.ref004 article-title: Structure of yellow fever virus envelope protein domain III publication-title: Virology doi: 10.1016/j.virol.2009.09.001 – start-page: 1043 volume-title: Fields Virology year: 2001 ident: pntd.0010770.ref006 – volume: 81 start-page: 825 issue: 5 year: 2009 ident: pntd.0010770.ref019 article-title: Dengue plaque reduction neutralization test (PRNT) in primary and secondary dengue virus infections: How alterations in assay conditions impact performance publication-title: Am J Trop Med Hyg doi: 10.4269/ajtmh.2009.08-0625 – ident: pntd.0010770.ref010 – volume: 11 start-page: e1001638 issue: 5 year: 2014 ident: pntd.0010770.ref009 article-title: Yellow Fever in Africa: estimating the burden of disease and impact of mass vaccination from outbreak and serological data publication-title: PLoS Med doi: 10.1371/journal.pmed.1001638 – volume: 38 start-page: 1823 issue: 5 year: 2000 ident: pntd.0010770.ref016 article-title: Standardization of immunoglobulin M capture enzyme-linked immunosorbent assays for routine diagnosis of arboviral infections publication-title: Journal of clinical microbiology doi: 10.1128/JCM.38.5.1823-1826.2000 – volume: 14 issue: 9 year: 2021 ident: pntd.0010770.ref005 article-title: The Present and Future of Yellow Fever Vaccines publication-title: Pharmaceuticals (Basel) doi: 10.3390/ph14090891 – start-page: 56 issue: 6 year: 2018 ident: pntd.0010770.ref025 article-title: Development of a Real-Time Reverse Transcription-PCR Assay for Global Differentiation of Yellow Fever Virus Vaccine-Related Adverse Events from Natural Infections publication-title: Journal of clinical microbiology – volume: 12 issue: 11 year: 2020 ident: pntd.0010770.ref008 article-title: Re-Emergence of Yellow Fever in Brazil during 2016–2019: Challenges, Lessons Learned, and Perspectives publication-title: Viruses doi: 10.3390/v12111233 |
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| Snippet | Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories chiefly... Background Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories... Molecular diagnosis of human specimens is the fastest way to confirm and subsequently mitigate yellow fever (YF) outbreaks. In Africa, a serological method... BackgroundEarly detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories... Background Early detection of human yellow fever (YF) infection in YF-endemic regions is critical to timely outbreak mitigation. African National Laboratories... |
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| SubjectTerms | Antibodies Assaying Biology and Life Sciences Cross-reactivity Detection Detection times Diagnosis Disease control Epidemics Evaluation Fever Humans Immunization International organizations Laboratories Mathematical analysis Medical laboratories Medicine and Health Sciences Microbiological strains Mitigation Molecular diagnostic techniques Nucleotide sequence Outbreaks PCR Performance evaluation Polymerase chain reaction Quality standards Research and Analysis Methods Reverse Transcriptase Polymerase Chain Reaction RNA Serology Serum Strains Strains (organisms) Tropical diseases Vaccines Vector-borne diseases Viruses Yellow fever Yellow Fever - epidemiology Yellow Fever Vaccine Yellow fever virus - genetics |
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| Title | Laboratory evaluation of RealStar Yellow Fever Virus RT-PCR kit 1.0 for potential use in the global yellow fever laboratory network |
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