Evaluation and validation of an RT-PCR assay for specific detection of monkeypox virus (MPXV)

Alberto Paniz-Mondolfi¹, Susana Guerra², Marina Muñoz³, Nicolas Luna³, Matthew M Hernandez¹, Luz H Patino¹, Jason Reidy¹, Radhika Banu¹, Paras Shrestha¹, Bernadette Liggayu¹, Audrey Umeaku¹, Feng Chen¹, Liyong Cao¹, Armi Patel¹, Ayman Hanna¹, Sunny Li¹, Andy Look¹, Nina Pagani^{4

5}, Randy Albrecht^{6

7}, Rebecca Pearl^{6

7}, Adolfo Garcia-Sastre^{1

6

7

8

9}, Dusan Bogunovic¹⁰, Gustavo Palacios^{1

6

7

8

9}, Lucia Bonnier¹¹, Freddy Cera¹¹, Heidi Lopez¹¹, Yvette Calderon¹², Erick Eiting¹², Karr Mullen¹², Sangyoon Jason Shin¹³, Luz Amarilis Lugo¹⁴, Antonio E Urbina¹⁴, Carlotta Starks¹⁵, Tonny Koo¹⁶, Patricia Uychiat¹⁷, Avery Look¹⁸, Harm van Bakel^{19

20}, Ana Gonzalez-Reiche^{19

20}, Adolfo Firpo Betancourt¹, David Reich¹, Carlos Cordon-Cardo¹, Viviana Simon^{1

6

7

8

21}, Emilia M Sordillo¹, Juan David Ramírez^{1

3}

Affiliations

¹ Department of Pathology, Molecular, and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
² Department of Preventive Medicine, Public Health and Microbiology, Universidad Autónoma de Madrid, Madrid, Spain.
³ Centro de Investigaciones en Microbiología y Biotecnología-UR (CIMBIUR), Facultad de Ciencias Naturales, Universidad del Rosario, Bogotá, Colombia.
⁴ Department of Biotechnology Laboratory Sciences, Valencia College, Orlando, Florida, USA.
⁵ Infectious Diseases Research Branch, Venezuelan Science Incubator and The Zoonosis and Emerging Pathogens Regional Collaborative Network, Cabudare, Lara, Venezuela.
⁶ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
⁷ Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
⁸ Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
⁹ The Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁰ Department of Microbiology, Center for Inborn Errors of Immunity, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹¹ Department of Pathology and Laboratory Medicine, Mount Sinai Beth Israel, New York, New York, USA.
¹² Department of Emergency Medicine, Mount Sinai Beth Israel, New York, New York, USA.
¹³ Department of Medicine, Ambulatory Care, The Center for Transgender Medicine and Surgery (CTMS) of Mount Sinai, Mount Sinai Beth Israel, New York, New York, USA.
¹⁴ Division of Infectious Diseases, Institute for Advanced Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁵ Mount Sinai Institute for Advanced Medicine, Jack Martin Fund Clinic and Comprehensive Health Clinic, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁶ Laboratory of Medical Microbiology, Mount Sinai Hospital, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁷ Department of Pathology and Laboratory Medicine, Mount Sinai West, New York, New York, USA.
¹⁸ Cornell University College of Agriculture and Life Sciences, Ithaca, New York, USA.
¹⁹ Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
²⁰ Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
²¹ Center for Vaccine Research and Pandemic Preparedness (C-VARPP), Icahn School of Medicine at Mount Sinai, New York, New York, USA.

PMID: 36271493
DOI: 10.1002/jmv.28247

Evaluation and validation of an RT-PCR assay for specific detection of monkeypox virus (MPXV)

Alberto Paniz-Mondolfi et al. J Med Virol. 2023 Jan.

. 2023 Jan;95(1):e28247.

doi: 10.1002/jmv.28247. Epub 2022 Nov 1.

Authors

Affiliations

¹ Department of Pathology, Molecular, and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
² Department of Preventive Medicine, Public Health and Microbiology, Universidad Autónoma de Madrid, Madrid, Spain.
³ Centro de Investigaciones en Microbiología y Biotecnología-UR (CIMBIUR), Facultad de Ciencias Naturales, Universidad del Rosario, Bogotá, Colombia.
⁴ Department of Biotechnology Laboratory Sciences, Valencia College, Orlando, Florida, USA.
⁵ Infectious Diseases Research Branch, Venezuelan Science Incubator and The Zoonosis and Emerging Pathogens Regional Collaborative Network, Cabudare, Lara, Venezuela.
⁶ Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
⁷ Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
⁸ Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
⁹ The Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁰ Department of Microbiology, Center for Inborn Errors of Immunity, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹¹ Department of Pathology and Laboratory Medicine, Mount Sinai Beth Israel, New York, New York, USA.
¹² Department of Emergency Medicine, Mount Sinai Beth Israel, New York, New York, USA.
¹³ Department of Medicine, Ambulatory Care, The Center for Transgender Medicine and Surgery (CTMS) of Mount Sinai, Mount Sinai Beth Israel, New York, New York, USA.
¹⁴ Division of Infectious Diseases, Institute for Advanced Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁵ Mount Sinai Institute for Advanced Medicine, Jack Martin Fund Clinic and Comprehensive Health Clinic, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁶ Laboratory of Medical Microbiology, Mount Sinai Hospital, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
¹⁷ Department of Pathology and Laboratory Medicine, Mount Sinai West, New York, New York, USA.
¹⁸ Cornell University College of Agriculture and Life Sciences, Ithaca, New York, USA.
¹⁹ Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
²⁰ Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
²¹ Center for Vaccine Research and Pandemic Preparedness (C-VARPP), Icahn School of Medicine at Mount Sinai, New York, New York, USA.

PMID: 36271493
DOI: 10.1002/jmv.28247

Abstract

Monkeypox virus (MPXV) is a zoonotic orthopoxvirus within the Poxviridae family. MPXV is endemic to Central and West Africa. However, the world is currently witnessing an international outbreak with no clear epidemiological links to travel or animal exposure and with ever-increasing numbers of reported cases worldwide. Here, we evaluated and validated a new, sensitive, and specific real-time PCR-assay for MPXV diagnosis in humans and compare the performance of this novel assay against a Food & Drug Administration-cleared pan-Orthopox RT-PCR assay. We determined specificity, sensitivity, and analytic performance of the PKamp™ Monkeypox Virus RT-PCR assay targeting the viral F3L-gene. In addition, we further evaluated MPXV-PCR-positive specimens by viral culture, electron microscopy, and viral inactivation assays. The limit of detection was established at 7.2 genome copies/reaction, and MPXV was successfully identified in 20 clinical specimens with 100% correlation against the reference method with 100% sensitivity and specificity. Our results demonstrated the validity of this rapid, robust, and reliable RT-PCR assay for specific and accurate diagnosis of MPXV infection in human specimens collected both as dry swabs and in viral transport media. This assay has been approved by NYS Department of Health for clinical use.

Keywords: diagnosis; monkeypox virus; orthopoxvirus; real-time PCR.

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References

REFERENCES

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Evaluation and validation of an RT-PCR assay for specific detection of monkeypox virus (MPXV)

Affiliations

Evaluation and validation of an RT-PCR assay for specific detection of monkeypox virus (MPXV)

Authors

Affiliations

Abstract

References

REFERENCES

MeSH terms

LinkOut - more resources

Full Text Sources

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Medical