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Monkeypox Virus (MPV) Nucleic Acid Detection Kit (Real-time PCR Method)
【Package Specification】
50 tests/kit & 200 tests/kit
【Intended Use】
This kit is used forthe detection of Monkeypox Virus(MPV) in serum or lesion exudate samples by using real
time PCR systems.
【Components of the Diagnostic Kit】
| No. | Reagent Name | Spec & Qty | |
| 50T | 200T | ||
| 1 | MPV PCR Mix | 750μl × 1 tube | 750μl × 4 tube |
| 2 | MPV Enzyme Mix | 200μl × 1 tube | 200μl × 4 tube |
| 3 | Internal control (IC) | 100μl × 1 tube | 100μl × 4 tube |
| 4 | MPV Negative Control | 100μl × 1 tube | 100μl × 4 tube |
| 5 | MPV Positive Control | 100μl × 1 tube | 100μl × 4 tube |
| 6 | Instructions | 1 serving | 1 serving |
【Test Method】
1. Reagent Preparation(performed at “reagent preparation region”)
1.1 Take out each components fromthe diagnostic kitand placethem at roomtemperature. Allowthe reagents to equilibrate at roomtemperature, then vortex each ofthem respectively forlater use.
1.2 Accordingto the quantity of test specimens, MPV Positive Control, MPV Negative Control ,and Internal control (IC),
pipette appropriate quantity of MPV PCR Mix and MPV Enzyme Mix ( MPV PCR Mix 15 μl/test +MPV Enzyme Mix 4 μl/test +Internal control (IC) 1 μl/test), mix themthoroughly to make a PCR-Mastermix, centrifugeit instantaneously forlater use.
| Reagent name | 1 Sample | 10 Sample | 25 Sample | 50 Sample | 100 Sample | 200 Sample |
| MPV PCR Mix(μl) | 15 | 150 | 375 | 750 | 1500 | 3000 |
| MPV Enzyme Mix(μl) | 4 | 40 | 100 | 200 | 400 | 800 |
| Internal control (IC) | 1 | 10 | 25 | 50 | 100 | 200 |
| PCR-Mastermix | 20 | 200 | 500 | 1000 | 2000 | 4000 |
| Note: The above configuration is just for your reference and to ensure enough volume of the PCR-Mastermix, more volume of the actual pipetting maybe required. | ||||||
1.3 Transferthe above-prepared reagents to the “specimen processing region” for later use.
2. Processing and loading of specimens (performed at “specimen processing region”)
2.1 This diagnostic kit does not include Viral RNA&DNA Extraction Kit. It is recommended to use Viral RNA&DNA Extraction Kitproduced by Changsha Renji Medical Equipment Co., Ltd. to extract viral DNA. The specific operation is in accordance with its instructions.
2.2 Add 20μl PCR-Mastermix into PCR reaction tube with 5μl above processed sample, MPV Positive Control and MPV Negative Control, and capthe tube. Carry out fluorescence quantitative PCR detection on fluorescence PCRinstrument.
3.PCR Amplification (performed at “nucleic acidamplification area”)
3.1 Place PCR reactiontubes into the specimen wells ofthe amplification equipment. Set up MPV Positive Control, MPV Negative Control and specimensto be tested in the corresponding sequence and input specimen name.
3.2 Set cycle parameters accordingto the followingtable for PCR amplification.
| Steps | Reaction Step | Temperature | Time | Cycles |
| 1 | UNG Enzyme Reaction | 50℃ | 2min | 1 |
| 2 | TAQ Enzyme Reaction | 95℃ | 3min | 1 |
| 3 | Deformation | 95℃ | 10s | 40 |
| Annealing | 55℃ | 30s* |
Note: 1)The fluorescence collection is set at "Step 3: 55 ° C, 30s". Selection of detection channels: FAM and Cy5, where FAM channel is Target Gene,Cy5 channel is Internal Control(IC) gene, and the reaction system is set to 25 μl.
2)ABI series fluorescent PCRinstruments do not select ROX calibration and select Noneforthe quenching group.