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Human cytomegalovirus nucleic acid quantitative detection primer and probe, kit and use thereof
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A technology of human cytomegalovirus and detection kit, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., and can solve problems such as false positives and cross-contamination
Active Publication Date: 2016-01-20
厦门安普利生物工程有限公司 +1
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However, since the amplified product needs to be detected by electrophoresis, it is easy to cause cross-contamination and false positive results
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[0101] 2. 8 samples to be tested are positive (Xiamen Hospital of Traditional Chinese Medicine) (P1, P2, P3, P4, P5, P6, P7, P8) (Clinically collect urine positive samples, the concentration is P11.0×10 7 copies / mL, P21.0×10 6 copies / mL, P31.0×10 5 copies / mL, P41.0×10 5 copies / mL, P51.0×10 4 copies / mL, P61.0×10 4 copies / mL, p71.0×10 3 copies / mL), p81.0×10 3 copies / mL).
[0102] ...
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Abstract
The invention provides a human cytomegalovirusnucleic acid quantitative detection primer and probe, a kit and application thereof. The primer has a sequence shown as SEQ ID NO:1 and SEQ ID NO:2; the probe has a sequence shown as SEQ ID NO:4, and a 5'-termination single strandbase sequence refers to 5-10 bases complementary to a target sequence and the middle 13-20 bases; a 3'-termination sequence refers to a sequence with 5-10 bases; the 5'-termination single strandbase sequence and a 3'-termination sequence are partially matched to form a neck structure; the probe has a neck ring structure; the TM value is equal to (neck ring TM value+7)+ / -1; 18-25 bases are arranged at the 5'-termination and can be completely matched with target genes, totaling about 23-32bp; fluorescent and non-fluorescent quenching groups are respectively labeled at the 5'-termination and 3'-termination. The primer and probe have a stable linear structure, are used for quantitatively detecting human cytomegalovirusnucleic acid and have the advantages of low fluorescent background value, high signal to noise ratio and high detection sensitivity.
Description
Technical field [0001] The present invention relates to in vitro diagnostic reagents. In particular, it relates to a primer and probe for quantitative detection of human cytomegalovirusnucleic acid, a kit and its use. Background technique [0002] Human Cytomegalovirus (HCMV) belongs to the herpes subfamily Group B of the Herpesvirus family. Existing human cytomegalovirus nucleic acid quantitative detection methods have a series of problems. For example, although electronmicroscopy technology is direct and objective, it requires high virustiter, expensive equipment, and requires skilled technicians, so the detection value is low. Virus isolation is not easy to carry out in large quantities due to the high technical requirements, long isolation time, and easy contamination. The detection of CMV-DNA by conventional PCR is simple and fast. However, since the amplified product needs to be detected by electrophoresis, it is easy to cause cross contamination and false positive re...
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