Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographies
ABSTRACT In the Global Polio Laboratory Network (GPLN), poliovirus (PV) screening results from acute flaccid paralysis (AFP) surveillance is based on virus isolation (VI) through cell culture, entailing long turnaround times and the amplification of live poliovirus. An alternative Direct Detection s...
Saved in:
| Main Authors: | , , , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2025-02-01
|
| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.02279-24 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832540832009289728 |
|---|---|
| author | Paula L. Marcet Brandon Short Ashley Deas Hong Sun Chelsea Harrington Shahzad Shaukat Muhammad Masroor Alam Marycelin Baba Adedayo Faneye Prossy Namuwulya Lea Necitas Apostol Tamer Elshaarawy John Kofi Odoom Peter Borus Shelina Moonsamy Yogolelo Riziki Marie Claire Endegue Zanga Mesfin Tefera Anfumbom K. W. Kfutwah Salmaan Sharif Varja Grabovac Cara C. Burns Nancy Gerloff |
| author_facet | Paula L. Marcet Brandon Short Ashley Deas Hong Sun Chelsea Harrington Shahzad Shaukat Muhammad Masroor Alam Marycelin Baba Adedayo Faneye Prossy Namuwulya Lea Necitas Apostol Tamer Elshaarawy John Kofi Odoom Peter Borus Shelina Moonsamy Yogolelo Riziki Marie Claire Endegue Zanga Mesfin Tefera Anfumbom K. W. Kfutwah Salmaan Sharif Varja Grabovac Cara C. Burns Nancy Gerloff |
| author_sort | Paula L. Marcet |
| collection | DOAJ |
| description | ABSTRACT In the Global Polio Laboratory Network (GPLN), poliovirus (PV) screening results from acute flaccid paralysis (AFP) surveillance is based on virus isolation (VI) through cell culture, entailing long turnaround times and the amplification of live poliovirus. An alternative Direct Detection strategy (DD-ITD) for screening viral nucleic acid from stools, bypassing the need for virus culture, has been developed and extensively validated by GPLN partners. A multi-laboratory demonstration project was conceived to field-test the DD-ITD method by GPLN laboratories from the WHO African, Western Pacific, and Eastern Mediterranean regions, where wild serotype 1 or vaccine-derived polioviruses still circulate. Strategically selected laboratories were tasked to simultaneously process stool suspensions with the current gold-standard VI method and the new DD-ITD strategy. Results from 12 laboratories were compiled and analyzed to assess the quality of each RNA extraction and rRT-PCR run. Matched results for both methods of over 10,500 specimens showed an overall method agreement of 91%. All laboratories detected more PV presumptive positive samples with the DD-ITD strategy than with VI, but a large proportion of DD-ITD positive results (72%) were inconclusive or non-typeable, requiring confirmation through sequencing. A total of 298 (2.8%) samples were PV positive using both methods, 828 (7.9%) positive only for DD-ITD, and 62 (0.6%) positive only with VI. The DD-ITD overall performance, quality of results, and agreement between method results varied significantly across participating laboratories. DD-ITD implementation would entail building proficiency in advanced molecular laboratory techniques and data analysis, and increased demand for confirmatory sequencing.IMPORTANCESurveillance of acute flaccid paralysis (AFP) and sensitive poliovirus detection are key components of the WHO Global Polio Eradication Strategy. This work summarizes the results of a multi-laboratory evaluation designed to field-test the performance and applicability of a molecular Direct Detection strategy (DD-ITD) that does not require amplification of live poliovirus. AFP samples were processed in parallel with both the DD-ITD and the current gold-standard PV detection methodology, based on virus isolation (VI) through cell culture. All participating laboratories detected more PV presumptive positive samples using the DD-ITD strategy than with virus isolation methodology, although a higher proportion of DD-ITD results required confirmatory sequencing. Significant variability among laboratories was observed in the quality of the results and overall DD-ITD performance. Implementing DD-ITD would entail building proficiency in advanced molecular laboratory techniques and strengthening data analysis skills. |
| format | Article |
| id | doaj-art-253a3b7e90194fa8ae32ac78172012a1 |
| institution | Kabale University |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-253a3b7e90194fa8ae32ac78172012a12025-02-04T14:03:41ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-02-0113210.1128/spectrum.02279-24Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographiesPaula L. Marcet0Brandon Short1Ashley Deas2Hong Sun3Chelsea Harrington4Shahzad Shaukat5Muhammad Masroor Alam6Marycelin Baba7Adedayo Faneye8Prossy Namuwulya9Lea Necitas Apostol10Tamer Elshaarawy11John Kofi Odoom12Peter Borus13Shelina Moonsamy14Yogolelo Riziki15Marie Claire Endegue Zanga16Mesfin Tefera17Anfumbom K. W. Kfutwah18Salmaan Sharif19Varja Grabovac20Cara C. Burns21Nancy Gerloff22Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USADivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USATanaq, Contracting agency to the Division of Viral Diseases, CDC, Atlanta, Georgia, USADivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USADivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USAWorld Health Organization, Global Polio Laboratory Network Headquarters, Geneva, SwitzerlandNational Institute for Health, Islamabad, PakistanNational Polio Laboratory, Maiduguri, NigeriaNational Polio Laboratory, Ibadan, NigeriaUganda Virus Research Institute, Entebbe, UgandaNational Polio Laboratory, Research Institute for Tropical Medicine, Manila, PhilippinesPolio Regional Reference Laboratory, VACSERA, Greater Cairo, EgyptNoguchi Memorial Institute for Medical Research, Accra, GhanaKenya Medical Research Institute, Nairobi, KenyaNational Institute of Communicable Diseases, Johannesburg, South AfricaNational Institute of Biomedical Research, Kinshasa, Democratic Republic of CongoCentre Pasteur du Cameroun, Yaoundé, CameroonNational Polio Laboratory, Addis Ababa, EthiopiaWorld Health Organization, Regional Office for Africa, Brazzaville, Democratic Republic of CongoWorld Health Organization, Regional Office for the Eastern Mediterranean, Amman, JordanWorld Health Organization, Regional Office for the Western Pacific, Manila, PhilippinesDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USADivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USAABSTRACT In the Global Polio Laboratory Network (GPLN), poliovirus (PV) screening results from acute flaccid paralysis (AFP) surveillance is based on virus isolation (VI) through cell culture, entailing long turnaround times and the amplification of live poliovirus. An alternative Direct Detection strategy (DD-ITD) for screening viral nucleic acid from stools, bypassing the need for virus culture, has been developed and extensively validated by GPLN partners. A multi-laboratory demonstration project was conceived to field-test the DD-ITD method by GPLN laboratories from the WHO African, Western Pacific, and Eastern Mediterranean regions, where wild serotype 1 or vaccine-derived polioviruses still circulate. Strategically selected laboratories were tasked to simultaneously process stool suspensions with the current gold-standard VI method and the new DD-ITD strategy. Results from 12 laboratories were compiled and analyzed to assess the quality of each RNA extraction and rRT-PCR run. Matched results for both methods of over 10,500 specimens showed an overall method agreement of 91%. All laboratories detected more PV presumptive positive samples with the DD-ITD strategy than with VI, but a large proportion of DD-ITD positive results (72%) were inconclusive or non-typeable, requiring confirmation through sequencing. A total of 298 (2.8%) samples were PV positive using both methods, 828 (7.9%) positive only for DD-ITD, and 62 (0.6%) positive only with VI. The DD-ITD overall performance, quality of results, and agreement between method results varied significantly across participating laboratories. DD-ITD implementation would entail building proficiency in advanced molecular laboratory techniques and data analysis, and increased demand for confirmatory sequencing.IMPORTANCESurveillance of acute flaccid paralysis (AFP) and sensitive poliovirus detection are key components of the WHO Global Polio Eradication Strategy. This work summarizes the results of a multi-laboratory evaluation designed to field-test the performance and applicability of a molecular Direct Detection strategy (DD-ITD) that does not require amplification of live poliovirus. AFP samples were processed in parallel with both the DD-ITD and the current gold-standard PV detection methodology, based on virus isolation (VI) through cell culture. All participating laboratories detected more PV presumptive positive samples using the DD-ITD strategy than with virus isolation methodology, although a higher proportion of DD-ITD results required confirmatory sequencing. Significant variability among laboratories was observed in the quality of the results and overall DD-ITD performance. Implementing DD-ITD would entail building proficiency in advanced molecular laboratory techniques and strengthening data analysis skills.https://journals.asm.org/doi/10.1128/spectrum.02279-24nucleic acid extractionpoliovirus eradicationacute flaccid paralysissurveillancedirect detectionreal-time RT-PCR |
| spellingShingle | Paula L. Marcet Brandon Short Ashley Deas Hong Sun Chelsea Harrington Shahzad Shaukat Muhammad Masroor Alam Marycelin Baba Adedayo Faneye Prossy Namuwulya Lea Necitas Apostol Tamer Elshaarawy John Kofi Odoom Peter Borus Shelina Moonsamy Yogolelo Riziki Marie Claire Endegue Zanga Mesfin Tefera Anfumbom K. W. Kfutwah Salmaan Sharif Varja Grabovac Cara C. Burns Nancy Gerloff Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographies Microbiology Spectrum nucleic acid extraction poliovirus eradication acute flaccid paralysis surveillance direct detection real-time RT-PCR |
| title | Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographies |
| title_full | Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographies |
| title_fullStr | Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographies |
| title_full_unstemmed | Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographies |
| title_short | Advancing poliovirus eradication: lessons learned from piloting direct molecular detection of polioviruses in high-risk and priority geographies |
| title_sort | advancing poliovirus eradication lessons learned from piloting direct molecular detection of polioviruses in high risk and priority geographies |
| topic | nucleic acid extraction poliovirus eradication acute flaccid paralysis surveillance direct detection real-time RT-PCR |
| url | https://journals.asm.org/doi/10.1128/spectrum.02279-24 |
| work_keys_str_mv | AT paulalmarcet advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT brandonshort advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT ashleydeas advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT hongsun advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT chelseaharrington advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT shahzadshaukat advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT muhammadmasrooralam advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT marycelinbaba advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT adedayofaneye advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT prossynamuwulya advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT leanecitasapostol advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT tamerelshaarawy advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT johnkofiodoom advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT peterborus advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT shelinamoonsamy advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT yogoleloriziki advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT marieclaireendeguezanga advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT mesfintefera advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT anfumbomkwkfutwah advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT salmaansharif advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT varjagrabovac advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT caracburns advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies AT nancygerloff advancingpolioviruseradicationlessonslearnedfrompilotingdirectmoleculardetectionofpoliovirusesinhighriskandprioritygeographies |