Monkeypox Virus Nucleic Acid Detection Kit (Real-time PCR)
[Specification] 50 tests/kit
This kit is used for in vitro qualitative detection of monkeypox virus nucleic acid in human rash fluid, nasopharyngeal swabs, throat swabs and serum samples。
Monkeypox (MP) is an acute zoonotic infectious disease caused by Monkeypox Virus (MPV). MPV is round-bricked or oval in shape, and is a double-stranded DNA virus with a length of about 197Kb. The disease is mainly transmitted by animals, and humans could become infected by
being bitten by infected animals or by direct contact with the blood, body fluids and rash of infected animals. The virus can also be transmitted between people, primarily through respiratory droplets during prolonged, direct face-to-face contact or through direct contact with a patient’s body fluids or contaminated objects[2-3]. The clinical symptoms of monkeypox infection in humans are similar
to those of smallpox, generally after a 12-day incubation period, appearing fever, headache, muscle and back pain, enlarged lymph nodes, fatigue and discomfort. The rash appears after 1-3 days of the fever, usually first on the face, but also in other parts. The disease course generally lasts 2-4 weeks, and the mortality rate is 1%-10%. Lymphadenopathy is one of the main differences between this disease and smallpox.
The test results of this kit are not used as the only indicator for the diagnosis of monkeypox virus infection in patients, which must be combined with the clinical characteristics of the patient and other laboratory test data to correctly determine the infection of the pathogen, and formulate a reasonable treatment plan to make the treatment safe and effective.
This kit adopts fluorescent probe real-time PCR technology, and select two specific conserved regions of monkeypox virus MPV-1 gene and MPV-2 gene to design specific primer probes. The MPV-1 gene probe 5’ is labeled with FAM, and the 3’ quenching group is labeled with BHQ1. The MPV-2 gene probe 5’ is labeled with VIC, and the 3’ quenching group is labeled with BHQ1. In the process of PCR amplification, specific primers and probes bind to their respective target sequences separately. When Taq enzyme encounters the probe bound to the target sequence, it exerts the function of 5’-end exonuclease, making the reporter fluorophore and the quenching group separated, thus the fluorescence monitoring system could receive the fluorescent signal, that is, a fluorescent molecule is formed every time a DNA chain is amplified, realizing that the accumulation of the fluorescent signal is completely synchronized with the formation of the PCR product. At the same time, endogenous internal control quality control is introduced to avoid false negatives in the test.
[Storage Conditions and Shelf Life]
The kit should be stored below -18℃ protected from light. The shelf life is 12 months. The number of repeated freezing and thawing should not be more than 4 cycles. It can be stably stored for 5 days below -18℃ and protected from light during transportation.
See the packaging label for the production date, batch number and expiration date.
Applied Biosystems™ Real time PCR system 7500, ABI 7500FAST real-time PCR system, ABI QuantStudio™5 Real-time PCR system, LightCycler® 480 fluorescence quantitative PCR system, Bio-Rad CFX96 real-time fluorescence quantitative PCR instrument, Ultrassay Archimed X4/X6 Real Time qPCR System. etc.
1.1 Rash Fluid
Select the typical or clear and fresh rash. Clean the surface twice with a sterile saline cotton swab and dipped the skin to dry, puncture the rash with a sterile needle tip, squeeze out with a sterile cotton swab and dip the rash fluid into the preservation solution immediately, and tighten the screw cap. Wipe the needle insertion site again with normal saline or sterile alcohol, and press with a sterile dry cotton swab for a while to prevent infection.
1.2 Nasopharyngeal swab
A swab with a polyester fiber tip is used to enter through the anterior nostril and proceed slowly back and forth along the base of the inferior meatus. When the top of the swab reaches the posterior wall of the nasopharyngeal cavity (with a touch of the wall), let the swab stand for a while (about 3 seconds), then gently rotate it for a round, slowly remove the swab, and put it into the tube. Break the plastic handle by hand, immerse the swab in the sampling solution, and tighten the cap. It is validated that nasopharyngeal swab samples can be preserved with normal saline or Hank’s solution preservation solution.
1.3 Oropharyngeal swab
Wipe the posterior pharyngeal wall and bilateral tonsils at the back of the uvula (palatine or uvula) with moderate force with a polyester fiber head swab. Avoid touching the tongue, take out the posterior sampling tube, and break the plastic handle at the contact part of the hand. Put the swab into the sampling solution, and tighten the cap. It is validated that oropharyngeal swab samples can be preserved with normal saline or Hank’s solution preservation solution.
Use a sterile syringe to collect 3-5mL of venous blood from the subject in a vacuum blood collection tube (preferably with a separating gel). Leave it at room temperature for no more than 4 hours, centrifuge at 3000 rpm for 10 minutes, suck the upper serum (do not suck red blood cells) and transfer it to another sterile centrifuge tube for use.