Description

Principle And Intended Use

This test is a qualitative real-time fluorescent PCR that enables to amplification of MPV from the oropharyngeal swab, nasopharyngeal swab, serum, plasma, and human pustular or vesicular rash specimens. Specific primers & probes are designed to detect the highly specific regions of the MPV. A pair of primers and a probe for detecting endogenous human genes are included as an internal control to monitor the whole test process and control for inhibition. The specific probe of the MPV gene is labeled with FAM, and the specific probe of the human gene is labeled with VIC.

The virus DNA in the sample should be purified and enriched with a nucleic acid extraction and purification reagent before being detected with this kit.

Specifications

96 tests/kit

Components

Each kit contains 1 pre-packaged reagent 96-well plate, 2 microplate sealers,1MPV Positive Control,

1 MPV Negative Control, and 1 instruction.

Storage Condition and Expiring Date
The reagents are stable when stored at room temperature for 12 months without unpacking. Please transport at -

25℃ to 35℃. The kit should remain sealed and away from light exposure. Do not use the kit beyond the expiration date.

Sample Result Judgment

① If the test sample detects a typical S-type amplification curve in the FAM channels and the Ct value is ≤35, and

there is a typical S-type amplification curve in the Endogenous Control channel (VIC), the sample can be

judged to be MPV positive.

② If the test sample has no amplification curve or Ct value> 38 in the FAM channels, and there is a typical

Stype amplification curve in the Endogenous Control channel (VIC), the sample can be judged to be MPV

negative.

③ If the test sample yields a Ct value ranging from 35-38, and there is a typical S-type amplification curve in the

Endogenous Control channel(VIC), the results need to retest. If the results repeated are consistent, and have a

typical S-type amplification curve, the sample can be judged positive, otherwise, the sample can be judged

negatively.

④ If no typical S-type amplification curve (No Ct value) or Ct value>38 is detected in the FAM and VIC channels of

the test sample, it means that there is a problem with the quality of the sample or a problem with the operation.

If the result is invalid, you should find and eliminate the cause, collect the sample again, and repeat the test(if

the test result is still invalid, please contact the company).

PCR Reaction Preparation & PCR Assay

PCR Reaction Preparation

① Take out the 96-well plate, equilibrate to room temperature (about 15-30 minutes), and cut off the required

strips based on the total number of samples (including patient specimen(s), MPV Positive Control, and MPV

Negative Control). PCR tube(s) can be cut down from the strip.

② Remove the microplate sealer on the plate, then add a 25µL sample (including extracted DNA from a patient

specimen(s), MPV Positive Control, and MPV Negative Control) to the PCR tube.

③ Cover the plate with the microplate sealers, mix, and transient centrifuge at 2000-6000 rpm, then put them into

the PCR instrument.

PCR Assay
① Create a new experiment.
② Select the FAM and VIC channels.
③ Select “none” for Passive Reference (For ABI Real-Time PCR systems).
④ Set cycle conditions. Enter the reaction volume (25μL), and edit the PCR program as follows:
Step 1：50℃ 2min；
Step 2：95℃ 5min；
Step 3：95℃ 10s，60℃ 40s (fluorescence collection ).（40 cycles）
Save the file. Run the program.

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Monkey Pox Virus qPCR Kit

Description

Sample Result Judgment

PCR Reaction Preparation & PCR Assay