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

被引：32

作者：

Paniz-Mondolfi, Alberto ^{[1
]}

Guerra, Susana ^{[2
]}

Munoz, Marina ^{[3
]}

Luna, Nicolas ^{[3
]}

Hernandez, Matthew M. ^{[1
]}

Patino, Luz H. ^{[1
]}

Reidy, Jason ^{[1
]}

Banu, Radhika ^{[1
]}

Shrestha, Paras ^{[1
]}

Liggayu, Bernadette ^{[1
]}

Umeaku, Audrey ^{[1
]}

Chen, Feng ^{[1
]}

Cao, Liyong ^{[1
]}

Patel, Armi ^{[1
]}

Hanna, Ayman ^{[1
]}

Li, Sunny ^{[1
]}

Look, Andy ^{[1
]}

Pagani, Nina ^{[4
,5
,6
]}

Albrecht, Randy ^{[7
,8
]}

Pearl, Rebecca ^{[7
,8
]}

Garcia-Sastre, Adolfo ^{[1
,7
,8
,9
,10
]}

Bogunovic, Dusan ^{[11
]}

Palacios, Gustavo ^{[1
,7
,8
,9
,10
]}

Bonnier, Lucia ^{[12
]}

Cera, Freddy ^{[12
]}

Lopez, Heidi ^{[12
]}

Calderon, Yvette ^{[13
]}

Eiting, Erick ^{[13
]}

Mullen, Karr ^{[13
]}

Shin, Sangyoon Jason ^{[14
]}

Lugo, Luz Amarilis ^{[15
]}

Urbina, Antonio E. ^{[15
]}

Starks, Carlotta ^{[16
,17
]}

Koo, Tonny ^{[18
]}

Uychiat, Patricia ^{[19
]}

Look, Avery ^{[20
]}

van Bakel, Harm ^{[21
,22
]}

Gonzalez-Reiche, Ana ^{[21
,22
]}

Betancourt, Adolfo Firpo ^{[1
]}

Reich, David ^{[1
]}

Cordon-Cardo, Carlos ^{[1
]}

Simon, Viviana ^{[1
,7
,8
,9
,23
]}

Sordillo, Emilia M. ^{[1
]}

David Ramirez, Juan ^{[1
,3
]}

机构：

[1] Icahn Sch Med Mt Sinai, Dept Pathol Mol & Cell Based Med, New York, NY 10029 USA

[2] Univ Autonoma Madrid, Dept Prevent Med Publ Hlth & Microbiol, Madrid, Spain

[3] Univ Rosario, Fac Ciencias Nat, Ctr Invest Microbiol & Biotecnol UR CIMBIUR, Bogota, Colombia

[4] Valencia Coll, Dept Biotechnol Lab Sci, Orlando, FL USA

[5] Venezuelan Sci Incubator, Infect Dis Res Branch, Cabudare, Lara, Venezuela

[6] Zoonosis & Emerging Pathogens Reg Collaborat Netw, Cabudare, Lara, Venezuela

[7] Icahn Sch Med Mt Sinai, Dept Microbiol, New York, NY 10029 USA

[8] Icahn Sch Med Mt Sinai, Global Hlth & Emerging Pathogens Inst, New York, NY 10029 USA

[9] Icahn Sch Med Mt Sinai, Dept Med, Div Infect Dis, New York, NY 10029 USA

[10] Icahn Sch Med Mt Sinai, Tisch Canc Inst, New York, NY 10029 USA

[11] Icahn Sch Med Mt Sinai, Ctr Inborn Errors Immun, Precis Immunol Inst, Dept Microbiol, New York, NY 10029 USA

[12] Mt Sinai Beth Israel, Dept Pathol & Lab Med, New York, NY USA

[13] Mt Sinai Beth Israel, Dept Emergency Med, New York, NY USA

[14] Mt Sinai Beth Israel, Dept Med, Ctr Transgender Med & Surg CTMS Mt Sinai, Ambulatory Care, New York, NY USA

[15] Icahn Sch Med Mt Sinai, Inst Adv Med, Div Infect Dis, New York, NY 10029 USA

[16] Icahn Sch Med Mt Sinai, Mt Sinai Inst Adv Med, Jack Martin Fund Clin, New York, NY 10029 USA

[17] Icahn Sch Med Mt Sinai, Comprehens Hlth Clin, New York, NY 10029 USA

[18] Icahn Sch Med Mt Sinai, Mt Sinai Hosp, Lab Med Microbiol, New York, NY 10029 USA

[19] Mt Sinai West, Dept Pathol & Lab Med, New York, NY USA

[20] Cornell Univ, Coll Agr & Life Sci, Ithaca, NY 14853 USA

[21] Icahn Sch Med Mt Sinai, Dept Genet & Genom Sci, New York, NY 10029 USA

[22] Icahn Sch Med Mt Sinai, Icahn Genom Inst, New York, NY 10029 USA

[23] Icahn Sch Med Mt Sinai, Ctr Vaccine Res & Pandem Preparedness, New York, NY 10029 USA

来源：

JOURNAL OF MEDICAL VIROLOGY | 2023年 / 95卷 / 01期

关键词：

diagnosis; monkeypox virus; orthopoxvirus; real-time PCR;

D O I：

10.1002/jmv.28247

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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 (TM) 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.

引用

页数：12

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