Diagnostic Performance of Mpox Virus (MPXV) Real-time PCR Assays: Multicenter Assessment and Extended Sensitivity Analysis

Overview

Journal Eur J Clin Microbiol Infect Dis

Publisher Springer

Specialties Infectious Diseases
Microbiology

Date 2024 Jun 4

PMID 38833104

Authors

Ziqiang Li

Yuqing Chen

Yanxi Han

Zhenli Diao

Tao Huang

Lei Feng

Yu Ma

Cong Liu

Meng Tian

Jing Li

Wanyu Feng

Zihong Zhao

Jian Jiang

Jinming Li

Rui Zhang

Affiliations

Soon will be listed here.

Abstract

Purpose: To comprehensively investigate the diagnostic performance of routinely used assays in MPXV testing, the National Center of Clinical Laboratories in China conducted a nationwide external quality assessment (EQA) scheme and an evaluated nine assays used by ≥ 5 laboratories in the EQA.

Methods: MPXV virus-like particles with 2700, 900 and 300 copies/mL were distributed to 195 EQA laboratories. For extended analysis, triple-diluted samples from 9000 to 4.12 copies/mL were repeated 20 times using the assays employed by ≥ 5 laboratories. The diagnostic performance was assessed by analyzing EQA data and calculating the limits of detection (LODs).

Results: The performance was competent in 87.69% (171/195) of the participants and 87.94% (175/199) of the datasets. The positive percentage agreements (PPAs) were greater than 99% for samples at 2700 and 900 copies/mL, and 95.60% (761/796) for samples at 300 copies/mL. The calculated LODs for the two clades ranged from 228.44 to 924.31 copies/mL and were greater than the LODs specified by the respective kits. EasyDiagnosis had the lowest calculated LODs and showed superior performance in EQA, whereas BioGerm and Sansure, with higher calculated LODs, did not perform well in EQA.

Conclusion: This study provides valuable information from the EQA data and evaluation of the diagnostic performance of MPXV detection assays. It also provided insights into reagent optimization and enabled prompt public health interventions for the outbreak.

References

Ladnyj I, Ziegler P, Kima E . A human infection caused by monkeypox virus in Basankusu Territory, Democratic Republic of the Congo. Bull World Health Organ. 1972; 46(5):593-7. PMC: 2480792. View

Mitja O, Ogoina D, Titanji B, Galvan C, Muyembe J, Marks M . Monkeypox. Lancet. 2022; 401(10370):60-74. PMC: 9671644. DOI: 10.1016/S0140-6736(22)02075-X. View

Aden D, Zaheer S, Kumar R, Ranga S . Monkeypox (Mpox) outbreak during COVID-19 pandemic-Past and the future. J Med Virol. 2023; 95(4):e28701. DOI: 10.1002/jmv.28701. View

Wang Y, Leng P, Zhou H . Global transmission of monkeypox virus-a potential threat under the COVID-19 pandemic. Front Immunol. 2023; 14:1174223. PMC: 10198437. DOI: 10.3389/fimmu.2023.1174223. View

Tarin-Vicente E, Alemany A, Agud-Dios M, Ubals M, Suner C, Anton A . Clinical presentation and virological assessment of confirmed human monkeypox virus cases in Spain: a prospective observational cohort study. Lancet. 2022; 400(10353):661-669. PMC: 9533900. DOI: 10.1016/S0140-6736(22)01436-2. View

Yin L, Man S, Ye S, Liu G, Ma L . CRISPR-Cas based virus detection: Recent advances and perspectives. Biosens Bioelectron. 2021; 193:113541. PMC: 8349459. DOI: 10.1016/j.bios.2021.113541. View

Lim C, Roberts J, Moso M, Liew K, Taouk M, Williams E . Mpox diagnostics: Review of current and emerging technologies. J Med Virol. 2022; 95(1):e28429. PMC: 10108241. DOI: 10.1002/jmv.28429. View

Adler H, Gould S, Hine P, Snell L, Wong W, Houlihan C . Clinical features and management of human monkeypox: a retrospective observational study in the UK. Lancet Infect Dis. 2022; 22(8):1153-1162. PMC: 9300470. DOI: 10.1016/S1473-3099(22)00228-6. View

Buchta C, Muller M, Griesmacher A . The importance of external quality assessment data in evaluating SARS-CoV-2 virus genome detection assays. Lancet Microbe. 2022; 3(3):e168. PMC: 8789236. DOI: 10.1016/S2666-5247(22)00003-9. View

10.

Zhang L, Sun Y, Chang L, Jia T, Wang G, Zhang R . A novel method to produce armored double-stranded DNA by encapsulation of MS2 viral capsids. Appl Microbiol Biotechnol. 2015; 99(17):7047-57. PMC: 7079959. DOI: 10.1007/s00253-015-6664-4. View

11.

Anderson M, Hodges A, Luk K, Olivo A, Forberg K, Meyer T . Development and Validation of Three Automated High-Throughput Molecular Tests to Detect Monkeypox Virus Infections. J Infect Dis. 2023; 229(Supplement_2):S137-S143. DOI: 10.1093/infdis/jiad406. View

12.

Armstrong G, MacCannell D, Taylor J, Carleton H, Neuhaus E, Bradbury R . Pathogen Genomics in Public Health. N Engl J Med. 2019; 381(26):2569-2580. PMC: 7008580. DOI: 10.1056/NEJMsr1813907. View

13.

Peeling R, Heymann D, Teo Y, Garcia P . Diagnostics for COVID-19: moving from pandemic response to control. Lancet. 2021; 399(10326):757-768. PMC: 8687671. DOI: 10.1016/S0140-6736(21)02346-1. View

14.

Islam M, Koirala D . Toward a next-generation diagnostic tool: A review on emerging isothermal nucleic acid amplification techniques for the detection of SARS-CoV-2 and other infectious viruses. Anal Chim Acta. 2022; 1209:339338. PMC: 8633689. DOI: 10.1016/j.aca.2021.339338. View

15.

Wei Z, Wang X, Feng H, Ji F, Bai D, Dong X . Isothermal nucleic acid amplification technology for rapid detection of virus. Crit Rev Biotechnol. 2022; 43(3):415-432. DOI: 10.1080/07388551.2022.2030295. View

16.

Shean R, Hymas W, Klein J, Pyne M, Hillyard D, Bradley B . Repeat testing of mpox specimens with late CTs improves detection of potential false positive cases. J Clin Microbiol. 2024; 62(2):e0127523. PMC: 10865812. DOI: 10.1128/jcm.01275-23. View

17.

Elnifro E, Ashshi A, Cooper R, Klapper P . Multiplex PCR: optimization and application in diagnostic virology. Clin Microbiol Rev. 2000; 13(4):559-70. PMC: 88949. DOI: 10.1128/CMR.13.4.559. View

18.

Deng W, Shi X, Tjian R, Lionnet T, Singer R . CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in fixed cells. Proc Natl Acad Sci U S A. 2015; 112(38):11870-5. PMC: 4586837. DOI: 10.1073/pnas.1515692112. View

19.

Ruijter J, Ramakers C, Hoogaars W, Karlen Y, Bakker O, van den Hoff M . Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res. 2009; 37(6):e45. PMC: 2665230. DOI: 10.1093/nar/gkp045. View