One-step Detection Kit for Hepatitis C Virus (QPCR-Probe)
[Packing Specification] 24 rxns/kit
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
purification. Test results should be combined with clinical
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.
HCV-PCR Reaction Mix: Specific premiers and fluorescent probe of
HCV, qRT-PCR Master mix.
Enzyme mix: Taq polymerase and Reverse transcriptase
HCV Positive Control: Pseudo virus with HCV gene established in
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
Analysis sensitivity: 40 copies/rxn
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
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
2)Separately add 2μL Positive control into the reactions as
3)Separately add 2μL Negative control to the reaction as negative
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||95ºC||5 second||35 cycles|
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.
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