Address:
Beijing，China
Main Products:
PCR thermal cycler,Real Time PCR Cycler,Dry Bath Incubator,One step gene test,Tissue Grinder
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FOB PriceNegotiable
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Min. Order1 Unit(s)
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Payment Terms-- --
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Port-- --
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Delivery Time5 days after payment
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Brand Name:Coyote
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Properties:Medical Materials &am...
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Place Of Origin:China
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Model Number:CRO 007V
[Product Name]
One-step Detection Kit for Hepatitis C Virus (QPCR-Probe)
[Packing Specification] 24 rxns/kit
[Intended Use]
This kit is used for the qualitative detection Hepatitis C Virus (HCV) in blood and serum samples. It offers auxiliary means for the diagnosis of the HCV infected patients without nucleic acid extraction and
purification. Test results should be combined with clinical diagnosis.
[Detection Principle]
This kit adopts PCR method combined with fluorescence probe in vitro amplification technology. In this method, The HCV probe contains a fluorescent reporter dye FAM at the 5' end of the probe and a quencher dye BHQ at the 3' end of the probe. The internal reference probe contains a fluorescent reporter dye VIC. When the probe is intact, the proximity of the reporter dye to the quencher dye suppresses the reporter fluorescence. Probe cleavage during the PCR reaction spatially separates the reporter dye from the quencher dye, thereby allowing detection of the reporter dye fluorescence. The fragments of reporter dye are displaced from the target, resulting in an increase in fluorescence. This step, which enables the fluorescence signal accumulation and PCR products formed synchronous, thus to achieve qualification detection the HCV in the infective patients' in serum samples, which provide auxiliary means for the HCV in the treatment of the patient.
[Kit Contents]
| Ref. | Type of reagent | 24 rxns |
| 1 | HCV-PCR Reaction Mix | 1 vial,1500μL |
| 2 | Enzyme mix | 1 vial,60μL |
| 3 | HCV-Internal control | 1 vial,60μL |
| 4 | HCV Positive Control | 1 vial,60μL |
| 5 | HCV Negative Control | 1 vial,200μL |
| 6 | Dilution Buffer | 1vial,1000μL |
| 7 | User Manual | 1 |
Enzyme mix: Taq polymerase and Reverse transcriptase
HCV Positive Control: Pseudo virus with HCV gene established in vitro, Internal recombinant plasmid
HCV Negative Control: RNase&DNase;-free water
Dilution Buffer: 1*PBS dilution buffer
Note: Different batches in the kit components should not be interchangeable
Analysis sensitivity: 40 copies/rxn
[Procedure]
1. Sample Preparation/Treatment
This kit does not need sample nucleic acid extraction and purification. Whole blood collected in sodium citrate anticoagulant. Serum sample should be separated from the whole blood for the following detection.
2. Preparation of amplification reagent
The Master Mix volume for each reaction should be pipetted as follow:
HCV-PCR mix 45 μL+ Enzyme mix 2μL+ HCV- Internal control 1μL Mix the components above before adding sample or controls
3.Adding samples and controls to the reaction tubes
Separately add the samples from step 1, positive control and blank control to different tubes:
1)Separately add 2 uL serum or 0.5 uL blood into the sample reaction tube.
2)Separately add 2μL Positive control into the reactions as positive control.
3)Separately add 2μL Negative control to the reaction as negative control.
Close the tube, mix thoroughly and spin down the mixture. Operation should be performed on the ice-bath. 2000rmp spin for 10sec. Put the reaction tube on the test instrument.
Note: to avoid the difference from the serum sample, it is recommend to use the serum sample which is clear. For the cloudy sample, if the first test result is negative, please dilute the sample to 4-10 folds using
the dilution buffer provided in the kit to detect again.
4.PCR Amplification
| Pro. | Tem. | Time | Cycle No. |
| 1 | 42ºC | 5min | 1cycle |
| 2 | 95ºC | 5 second | 15cycles |
| 45ºC | 5 second | ||
| 3 | 95ºC | 1min | 1cycle |
| 4 | 95ºC | 5 second | 35 cycles |
| 50ºC | 10 second (acquire fluorescence) |
Selection of fluorescence channels: 530nm channel(Reporter FAM): HCV; 560nm channel(Reporter VIC): internal control; Please refer to the instrument manual for specific channel set detection.
[Results interpretation]
a. if the sample Ct value ≤28, report positive HCV;
b. if the sample Ct value 30>Ct>28,Please retest the sample. If the repeat result still>28 and the negative control is no value, the result is positive sample, and if the repeat sample result is no value, report as HCV negative.
c. if the sample Ct value is >28 or no value, report negative for HCV.
