quanty Monkeypox

The quanty Monkeypox system is a quantitative test that allows the DNA amplification and quantification, by means of Real-Time PCR, of the 2022 Monkeypox virus genome.

Overview

Monkeypox virus is an enveloped double-stranded DNA virus that belongs to the Orthopoxvirus genus of the Poxviridae family. There are two distinct genetic clades of the monkeypox virus: the central African (Congo Basin) clade and the west African clade. The 2022 MPXV sequences from Europe are part of a distinct cluster within the West African clade and cluster together with 2022 MPXV sequences from Brazil, Canada and the US. Monkeypox is usually a self-limited disease with the symptoms lasting from 2 to 4 weeks. Severe cases occur more commonly among young person and are related to the extent of virus exposure, patient health status and nature of complications. Underlying immune deficiencies may lead to worse outcomes. Although vaccination against smallpox was protective in the past, today persons younger than 40 to 50 years of age (depending on the country) may be more susceptible to monkeypox due to cessation of smallpox vaccination campaigns globally after eradication of the disease.  Complications of monkeypox can include secondary infections, bronchopneumonia, sepsis, encephalitis, and infection of the cornea with ensuing loss of vision. Detection of viral DNA by Real-Time PCR is the preferred laboratory test for monkeypox given its accuracy and sensitivity.

European Centre for Disease Prevention and Control/WHO Regional Office for Europe. Monkeypox, Joint Epidemiological overview, 30 June, 2022.

The quanty Monkeypox kit can detect the MPXV genome and also discriminate the two MPXV clades (West Africa and Congo basin).

Target:

• J2L/J2R common region of MPXV genome for the screening (MPXV)
• J2L/J2R specific region of west African MPXV genome (MPXV-WA)
• D14L specific region of Congo basin MPXV genome (MPXV-CB)
• Internal Control (IC)

Standard for quantification:

• Synthetic DNA corresponding to the J2L/J2R screening region  (100.000 cps/µL - 100 cps/µL)

Diagnostic samples:

• Biological samples (for example swabs from skin lesions)

Real-Time PCR instruments:

• Applied Biosystems 7500 Fast (ThermoFisher SCIENTIFIC)
• Rotor-Gene Q MDx (RG-Q MDx - QIAGEN)
• CFX96 Real-Time PCR Detection System (Bio-Rad)

DNA Extraction:

Manual Extraction with:

• QIAmp DNA mini kit  (QIAGEN)
• Bact Extra Pure Kit (ref. EDR004050).

Automatic Extraction with:

• CloNext 12 or CloNext 24 (ref. OP01018/OP01019). Robotic Workstation for the automatic purification of the nucleic acids 
• Duplicα®Prep (ref. EDI001 or EDI003). Robotic Workstation for the automatic purification of the nucleic acids