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HWTS-HP009A HCV RNA Quantitative Detection Kit(Fluorescence PCR)

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Category:Hepatitis

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HWTS-HP009A HCV RNA Quantitative Detection Kit(Fluorescence PCR)

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Introduction

[REF] HWTS-HP009A

[Specification] 50 tests/kit

[Research Use Only]

Hepatitis C Virus (HCV) nucleic acids in human blood plasma or serum samples with an aid of Quantitative Real-Time Polymerase Chain Reaction (qPCR) method.

The hepatitis C virus (HCV) is a small, enveloped, single-stranded, positive-sense RNA virus. It is a member of the genus Hepacivirus in the family Flaviviridae. HCV is spread primarily by direct contact with human blood. It is a major cause of acute hepatitis and chronic liver disease, including cirrhosis and liver cancer.

[Test Principle]

This test kit is a quantitative real-time RT-polymerase chain reaction assay for detecting hepatitis C virus (HCV) RNA in human plasma and serum. Nucleic acids of HCV were extracted from plasma/serum using spin column. The conserved region of hepatitis C virus was selected as the detection target gene. The specific primers and fluorescent probe were designed. The fluorophore is attached at the 5' end of the probe and the quencher moiety is located at the 3' end. A separate probe with VIC fluorophore at the 5’ end is set as internal control in the assay to rule out the failure of amplification in cases where the target sequence is not detected. The extracted RNA was reversely transcribed into cDNA, and the fluorescence signal was detected by fluorescence quantitative PCR The quantitative detection of nucleic acid in samples can be realized by the change of fluorescent signal value of RNA standard amplification; the processing process of samples to be tested and the existence of PCR inhibition reaction can be monitored by detecting whether the amplification of internal control is normal, so as to avoid false negative detection.

 

[Storage Conditions and Shelf Life]

HCV RNA quantitative kit should be stored in dark at temperature lower than -18°C stable with self-life about 9 months. Repeated freezing-thawing cycles (5 x) should be avoided.

[Applicable Equipment]

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 XP96 Real Time qPCR System. etc.

[Acceptable Specimens]

Specimen collection

Human blood plasma and serum are the most suitable sample materials for HCV detection. 2mL venous blood should be withdraw on the appropriate parts of the elbow bend, the forearm, or the back of the hand using sterile syringe and collected in standard specimen collection tubes or anticoagulation tubes (EDTA·K2 or Sodium citrate).

The whole blood should be centrifuged at 1600 rpm for 20 minutes within 4 hours to separate serum or plasma. The separated serum or plasma must be transferred to a sterile centrifuge tube. Be sure not to draw red cell in transferring.

Blood specimen in tube with Heparin sodium as anticoagulant cannot be used in the assay.

Storage The specimen should be kept at 2~8°C no longer than 1 week, -18°C for no longer than 6 months and -70°C for longer term preservation. Repeated freezing-thawing should be avoided.

Transportation

Specimen should be transported frozen using sealed container with ice bag or drikold.

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