As one of the leading companies in Europe, PentaBase has developed and produced a qPCR assay, specific and sensitive for the analysis of Monkeypox, validated on Monkeypox DNA extracted from Danish citizens by Rigshospitalet.
ASSAYS | Monkeypox
ASSAYS | Monkeypox
Introduction to InfectionDetect™ Monkeypox
Since early May 2022, an increase in Monkeypox cases has been reported from non-endemic countries. The virus has spread from West and Central Africa to North America and Europe. There have never been reported such a significant number of cases before. WHO is trying to prevent the further spread of the disease by guidance and increased surveillance¹. Once again, PentaBase has responded quickly to a new outbreak, and designed and produced a qPCR assay, specific and sensitive for the analysis of Monkeypox, validated on Monkeypox DNA extracted from Danish citizens by Rigshospitalet (Research-Use-Only).
Infection Detect™ Mpox qPCR Assay benefits:
2 Viral Sequences
Multiplex Assay
LOD5
Developed in collaboration with Rigshospitalet
Detailed information
Two Viral Genes
InfectionDetect-Mpox qPCR assay from PentaBase is targeting two Monkeypox genome DNA sequences, J2R (HEX) and B7R (FAM). The two Monkeypox-specific regions have been validated in-silico against other orthopox viruses and have been found to be specific. The assay comprises positive and negative controls. The positive control contains two synthesized sequences of the monkeypox virus with a length of 300 base pairs. The two sequences are not connected with pathogenicity and the Centre for Biosecurity and Biopreparedness (CBB) has declared them not subject to control following Danish legislation (BEK no. 981 of 15/10/2009).
Internal Control
The assay uses the human RNAse P (Cy5) as a swabbing and amplification control. The assay is for research use only. The positive control contains two synthesized sequences of the monkeypox virus with a length of 300 base pairs.
Validation
PentaBase systems for analysis
The analysis is developed on the BaseTyper™ real-time PCR instruments but will be validated on Bio-Rad’s CFX, Thermofishers’ Quant-Studio5 and Roche’s LightCycler480. Furthermore, the assay will be able to run on the fully automatic Cobas 6800 and 8800 in an open channel. The instrument time required for the analysis is 53 minutes when using a BaseTyper™.
For purification, we recommend the Viral RNA and DNA purification kit for the BasePurifier™, but other viral purification systems such as one from Biomerieux have also shown to work.
Sampling can be done using individual swabs for separate anatomic sites (rectal, oropharyngeal or skin lesions) added in individual transportation tubes comprising inactivating transport media (ITM) for the safety of downstream handling.
