Title : Clinical evaluation of Mpox virus detection by real time PCR
Abstract:
Background: In an increasingly globalized and interconnected world, the outbreak of an infectious disease in one country can become a worrying health emergency for the whole world. A recent example is the 2022 Mpox outbreak affecting multiple areas across the world. In this context, Global health systems need to develop effective strategies to interrupt transmission as soon as possible by identifying cases, clusters, and sources of infection. The aim of this retrospective and collaborative study was to perform the external clinical validation of a ready-to-use reagent product designed by Certest Biotec for the detection of Mpox virus by qPCR.
Methods: A total of 165 samples suspected of Mpox virus were used for this analysis. The standard procedure of the Microbiology Laboratory of the Miguel Servet University Hospital was considered as reference techniques (RealStar Orthopoxvirus PCR kit 1.0, Altona Diagnostics and bidirectional sanger sequencing). The use of all data and samples was approved by the research ethics committee of Aragon (CEICA) (PI22/412; 5 Oct 2022).
Results: Accuracy tests showed good clinical validation values (Table 1): Sensitivity 1 (0.97-1); Specificity 1 (0.98-1); Positive predictive value 1 (0.93-1) and Negative predictive value 1 (0.95-1). Strength of agreement was almost perfect (K=1; P<0.001) and spearman correlation test yielded strong correlation (rs=0.78; P<0.001). All positive samples were sequenced and all generated sequences with suitable sizes were blasted for species classification confirming Mpox virus presence.
|
TP |
TN |
FP |
FN |
Sensitivity |
Specificity |
PPV |
NPV |
K |
|
69 |
96 |
0 |
0 |
1 (0.97-1) |
1 (0.98-1) |
1 (0.93-1) |
1 (0.95-1) |
1 |
Conclusions: This retrospective study demonstrates the good clinical parameters and the strong overall agreement between the VIASURE® Monkeypox virus Real Time PCR detection kit (CerTest Biotec) compared to both the RealStar Orthopoxvirus PCR kit 1.0 and the Sanger sequencing results. The added value observed is the useful support to clinicians in their specific diagnosis of Mpox virus infections due to the diagnostic specificity data obtained.
Furthermore, the stabilized and ready-to-use format allows storage at room temperature, enabling a longer shelf life. This is an important advantage in laboratories where refrigerated storage space is limited. It also facilitates transport, so there is no need for special cold packaging, making it environmentally safe and reducing additional costs.
Audience take away:
- A new tool for molecular diagnosis of Mpox virus is presented in a stabilized and ready-to-use format.
- The format of the test allows storage at room temperature and a longer shelf life. This is an important advantage in laboratories where refrigerated storage space is limited. It also facilitates transport, so there is no need for special cold packaging, making it environmentally safe and reducing additional costs
- The clinical performance of the test is equivalent to reference methods currently used in health systems for Mpox virus detection.
