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

Alberto Paniz-Mondolfi, Susana Guerra, Marina Muñoz, Nicolas Luna, Matthew M. Hernandez, Luz H. Patino, Jason Reidy, Radhika Banu, Paras Shrestha, Bernadette Liggayu, Audrey Umeaku, Feng Chen, Liyong Cao, Armi Patel, Ayman Hanna, Sunny Li, Andy Look, Nina Pagani, Randy Albrecht, Rebecca PearlAdolfo Garcia-Sastre, Dusan Bogunovic, Gustavo Palacios, Lucia Bonnier, Freddy Cera, Heidi Lopez, Yvette Calderon, Erick Eiting, Karr Mullen, Sangyoon Jason Shin, Luz Amarilis Lugo, Antonio E. Urbina, Carlotta Starks, Tonny Koo, Patricia Uychiat, Avery Look, Harm van Bakel, Ana Gonzalez-Reiche, Adolfo Firpo Betancourt, David Reich, Carlos Cordon-Cardo, Viviana Simon, Emilia M. Sordillo, Juan David Ramírez

Research output: Contribution to journalResearch Articlepeer-review

25 Scopus citations

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.

Original languageEnglish (US)
Article numbere28247
JournalJournal of Medical Virology
Volume95
Issue number1
DOIs
StatePublished - Oct 19 2023

All Science Journal Classification (ASJC) codes

  • Infectious Diseases
  • Virology

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