Monkeypox virus

Molecular detection of Monkeypox virus nucleic acid
Microbiology
Virology
○ Lesion fluid, crust material or scab ○ Lesion swab ○ Nasopharyngeal and/or throat swab ○ Serum ○ CSF ○ Urine ○ Frozen tissues/formalin-fixed or paraffin embedded tissues
Other Information
○ NP and throat swabs are particularly useful specimens if testing during the prodrome (e.g. pre-rash in febrile patients who are contacts of cases, patients with macular or papular rash) – and may be collected on all patients.
○ Submit a maximum of 3 skin lesion specimens per patient.
○ NP/throat swabs and blood specimens are generally not recommended in patients who have skin lesions that can be swabbed, or have skin lesion material that can be submitted (e.g. scab). Skin lesions usually have higher viral loads, therefore skin swabs or lesion material are more sensitive (90%) for monkeypox virus detection. However, blood should always be submitted along with a NP swab or throat swab on patients suspected of mpox infection who do not have a skin rash or their skin rash can’t be reliably swabbed.
○ Anal or rectal swabs are recommended on patients with lesions in these locations or symptoms of involvement e.g. rectal pain.
○ All specimens from a patient being investigated for mpox, including specimens submitted for other tests, should indicate on the requisition that this patient is suspected of having mpox.
○ Lesion fluid and/or crust/tissue: Sterile container
○ Lesion swab/NP swabs/throat swabs: Swab in viral transport medium
○ Serum: red top or serum separator tube (clotted tube) (0.5 mL)
○ CSF (0.5mL), urine (50 mL): Sterile container
○ Frozen tissues/paraffin embedded tissues: entire blocks or four to six 10-micro block sections

Lesion fluid and/or crust/tissue:
○ Collect lesion or crust material and scabs in sterile container

Lesion swab:
○ Collect lesion swab by cleaning vesicle with saline, disrupting vesicle, and collecting fluid and cells from lesion base. If nonvesicular, collect cells from lesion base with a swab premoistened with saline.

Nasopharyngeal swab
○ Insert tip of swab into nostril aiming along the floor of nasal cavity until posterior wall of nasopharynx is reached. Resistance will be felt when reached. Rotate swab for 10-30s. Place swab into transport medium and close tube. Repeat collection with second nostril.

Throat swab:
○ Collect the specimen using a clean, sterile swab by swabbing the posterior oropharynx vigorously and placing it in universal transport medium.

Serum:
○ Decontaminate skin with 70% alcohol followed by iodine or chlorhexidine, collect at least 5 mL of blood.

Urine:
○ Collect urine into a clean, sterile container. Transport to the microbiology laboratory ASAP.

CSF:
○ Decontaminate skin, collect at least 0.5 mL of CSF by lumbar puncture.

Tissue
Paraffin-embedded or formalin fixed tissues can be sent to the laboratory labelled as such in a sterile container.

Refer to section Sample Processing / Delivery
PHOL
○ Up to 2 days from receipt by PHOL
416-586-4800 extension 4559
NO
○ Label specimen container. Place specimens in biohazard bag and seal.

○ Specimens should be stored and transported to the UHN/SH Microbiology Laboratory at refrigeration temperature (2-8˚C) immediately after collection.

○ The specimen will be referred to the Public Health Ontario Laboratory for testing and will be referred to NML.