Method for semi-quantitatively detecting pathogenic nucleic acid by adding internal control nucleic acid
An internal control nucleic acid and semi-quantitative technology, which is applied in the field of semi-quantitative detection of pathogen nucleic acid by adding internal control nucleic acid, can solve the problems of false negatives, achieve the effect of convenient operation, low production cost, and easy promotion
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Embodiment 1
[0047] Embodiment 1 (adopt the test strip method that adds internal control, detect HBV virus in the blood)
[0048] my country is a high-incidence area of hepatitis B virus infection in the world. At present, hundreds of millions of people in the country are carriers of hepatitis B virus, and there are about tens of millions of chronic hepatitis patients. Every year, the cost of direct treatment of viral hepatitis in my country is about tens of billions (excluding the cost of health care products, the indirect losses caused by affecting studies, employment and work, etc.). Efficient and accurate detection of HBV is crucial in its prevention and treatment.
[0049] The HBV genome is a circular double-stranded DNA with a full length of 3182 bp (about 3.2 kb). The length of the double strand is asymmetric, and the negative strand (long strand) is complementary to the viral mRNA. Only the 5' end of the positive strand is fixed, and its length can be 50%-100% of the negative s...
Embodiment 2
[0063] Embodiment 2 (investigate the semi-quantitative performance of the test strip method adding internal control)
[0064] experiment process:
[0065] 1) PCR amplification: Six different concentrations of plasmid templates (5×10 1 , 5×10 2 , 5×10 3 , 5×10 4 , 5×10 5 , 5×10 6 copies / μL) and a certain concentration of HBV internal control nucleic acid fragments (50copies / μL) were mixed as templates, and HBV sample amplification-specific primers and corresponding internal control primers were used for double amplification in a single tube. The HBV internal control nucleic acid fragment sequence, the HBV primer sequence, the corresponding internal control nucleic acid fragment primer sequence, the reaction system and the reaction procedure are the same as in Example 1.
[0066] 2) Test strip detection: the implementation steps are the same as above, see the results Figure 5 . Depend on Figure 5 It can be seen from the results that when the concentration of the viral...
Embodiment 3
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