Human Papillomavirus (high risk subtype) HPV test Kit (PCR method) luorescent PCR Kit for Human Papilloma Virus (HPV) Genotyping

Product Description

Quick-Dot Human Papillomavirus HPV test Kit (PCR method)

Name: Human papillomavirus (high risk subtype) nucleide detection system kit (PCR-fluorescent probe method)

Specification: *4 pieces/ box or *8 pieces/ box

Type: the kits are divided into Type I and Type II:

a)       Type I can detect whether its single or compound HPV high risk subtype infection, but it cant confirm which type or types it is or they are.

b)       Type II can detect whether its single or compound HPV high risk subtype infection, and it can judge whether its Type *6 or Type *8.

Expected purpose: Its used to qualitatively detect *4 high risk subtypes (HPV*6,*8,*1,*3,*5,*9,*5,*1,*2,*6,*8,*9,*6,*8) of human papillomavirus DNA in womens cervical cells.

Principle of test: Based on human papillomavirus DNA as the template, it takes use of PCR fluorescent method to detect the infection condition.

Major composition:

Storage condition and validity: it should be preserved under temperature of **8C~ **0C, kept in darkness. The validity is *2 months , please use the kits in validity.

Applicable instruments: ABI***0, Bio-Rad CFX*6, Roche LC**0 etc.

Specimen requirements:

1, Preparation before test:

1)       Menstruation is normal and the test should be taken ****8 days after menses for the best results.

2)       No sexual behavior in *4 hours before test.

3)       No vinegar or iodine solution smeared before test.

4)       No drug used or washed in the vagina in *2 hours before test , or itll influence the test results.

2, Specimen collection method and steps:

1)       Gently wipe up the excessive cervical secretion with cotton swabs. The cytobrush should be close to the cervical mucosa. And it should be clockwise rotated for **6 turns to collect the enough deciduous cervical cells.

P.s. Please strictly follow the above specimen collection steps, the quantity of the deciduous cervical cells will directly affect the accuracy of testing results.

2) Carefully take out the cytobrush and put it into specimen saving tube which is equipped with 3ml sterile Isotonic phosphate buffer. The deciduous cervical cells attached with the cytobrush should be wholly washed into the tube. Then the hair of cytobrush should be well wiped and thrown away. The tube cap should be screwed down, the tube should be labeled with patients number, and then be delivered to inspection.

Testing method:

1, DNA extraction:

Take 1ml sterile Isotonic phosphate buffer with deciduous cervical cells out (if the quantity of cells is small, the volume of the liquid should become 2ml), and transfer the liquid into a new centrifugal tube.

Put the tube into a centrifugal machine, rotated with a speed of *0,**0~ *3,**0    turns/second, to be centrifuged for *0 mins, and then abandon the liquidsupernatant. The deciduous cervical cells will form sediment on the bottom of the tube(the amount of the sediment doesnt indicate the quantity of deciduous cervical cells).

Add *0ul Virus cracking liquid into the tube, shake and mix it up , heat the tube up till boiled for *0 mins. Resuspend the deciduous cervical cells. The cells should be sufficiently resuspended by a Whirlpool blending oscillator so that the quantity of DNA will be more.

Put the compound liquid of step 3 in a centrifugal machine, rotated in a speed of *0,**0 ~ *3,**0 turns/ min, to be centrifuged for *0 mins. The liquidsupernatant is regarded as the template of PCR reaction now.

Take 2ul PCR reaction template and the rest is preserved under temperature of **0c.

2, Add specimen into DNA template

Carefully add 2ul PCR reaction template into PCR reaction tube. We suggest you should set up negative and positive contol groups for each test. You should add 2ul positive and negative quality control liquid into relative PCR tubes.

3, Preparation of amplification reagents:

Unfreeze the reagents. Take out the Human papillomavirus (high risk subtype) nucleide detection system kit and unfreeze it until it returns to the room temperature.

Centrifuge the reagents. Make sure the reagents attached with the cap and tube have been centrifuged to the bottom of the tube and mixed up.

Allocate the PCR reaction premix compound liquid as the following table required. And then put the liquid in the centrifugal machine and rotate it, make sure the eagents attached with the cap and tube have been centrifuged to the bottom of the tube and mixed up.(due to the loss caused by adding the specimen, we suggest that you should prepare one more piece of premix liquid for every *0 pieces.)

Allocate the PCR reaction premix liquid into the reaction holes of the existing templates as *3ul per capita. And seal the fluorescent qualitative PCR level sealing film or screw up the cap and rotate with a speed of 2,**0 turns/min, centrifuge for *0s. And make sure the liquid has been centrifuged into the bottom.

4, PCR reaction procedure

Set up the reaction conditions as following table required based on your PCR instrument.