Primer probe combination and kit for early diagnosis, metastasis early warning and prognosis evaluation of prostate cancer and application of primer probe combination and kit

A technology for early diagnosis of prostate cancer, applied in the direction of recombinant DNA technology, biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of complex detection process and high risk of contamination, and achieve high PCR efficiency, good stability, highly specific effect

Pending Publication Date: 2022-04-29
SHANGHAI CONLIGHT MEDICAL LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] The purpose of the present invention is to provide a combination of primers and probes, a kit and its application for early diagnosis of prostate cancer, early warning of metastasis and assessment of prognosis, so as to solve the problem of complex detection process and high risk of contamination in the existing early diagnosis technology of prostate cancer At the same time, the existing technology lacks the problem of metastasis early warning and prognosis assessment for prostate cancer

Method used

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  • Primer probe combination and kit for early diagnosis, metastasis early warning and prognosis evaluation of prostate cancer and application of primer probe combination and kit
  • Primer probe combination and kit for early diagnosis, metastasis early warning and prognosis evaluation of prostate cancer and application of primer probe combination and kit

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Embodiment 1

[0035] The design of embodiment 1 primer, probe

[0036] In this example, the inventors designed primers and probes for detecting PCA3 mRNA, PSA mRNA, TMPRSS2-ERG, and ACTB genes respectively according to the sequence characteristics of PCA3 mRNA, PSA mRNA, TMPRSS2-ERG, and ACTB genes. The specific sequences of the needles are shown in Table 1 below.

[0037] Table 1

[0038] Primer probe name Sequence and Modification PCA3-F (SEQ ID NO: 1) AAAGGAAGCACAGAGATCCCCTG PCA3-R (SEQ ID NO: 2) GGGCGAGGCTCATCGAT PCA3-P (SEQ ID NO: 3) FAM-AGAAATGCCCGGCCGCCATC-BHQ1 PSA-F (SEQ ID NO: 4) GACCACCTGCTACGCCTCA PSA-R (SEQ ID NO: 5) GGAGGTCCACACACTGAAGTTTC PSA-P (SEQ ID NO: 6) HEX-CAGCATTGAACCAGAGGAGTTCTTGACCC-BHQ1 TMPR-ERG-F (SEQ ID NO: 7) GCCGCCTGGAGCGCGGCA TMPR-ERG-R (SEQ ID NO: 8) CTGCCGCACATGGTCTGTACTCC TMPR-ERG-P (SEQ ID NO: 9) ROX-ACTCTGCGCTCGTTCGTGGTCAT-BHQ2 ACTB-F (SEQ ID NO: 10) GGCACCCAGCACAATGAAG ...

Embodiment 2

[0040] The extraction of embodiment 2 nucleic acid samples

[0041] 2.1 Urine exosome extraction

[0042] 1) Collect the urine sample, centrifuge at 3000×g for 30 min at 2-8°C, and collect the supernatant to remove residual cells and debris in the sample;

[0043] 2) Take 0.9mL of the above sample, add 0.3mL EPS-U solution, invert and mix well, and let stand overnight at 2-8°C;

[0044] 3) Centrifuge the sample at 12000×g for 30 minutes in a centrifuge, and discard the supernatant;

[0045] 4) Resuspend the pellet with 100-200 μL ERS-U and store at 2-8°C or perform nucleic acid extraction directly.

[0046] 2.2 Extraction of nucleic acid samples

[0047] 1) The preparation and inspection of reagent materials before the experiment are as follows:

[0048] Check the shelf life of the nucleic acid kit (kit name) and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding marks on the bottle; (2) isopropanol (if not available, absolute ethanol can...

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Abstract

The invention provides a primer probe combination for early diagnosis, metastasis early warning and prognosis evaluation of prostate cancer, a kit and application of the primer probe combination and the kit. The primer probe combination comprises a first group of primer pairs, a second group of primer pairs and a third group of primer pairs, wherein the first group of primer pairs is used for detecting the PCA3 gene and has nucleotide sequences as shown in SEQ ID NO: 1-2; the second group of primer pairs is used for detecting the PSA gene and has nucleotide sequences as shown in SEQ ID NO: 4-5; the third group of primer pairs is used for detecting the TMPRSS2-ERG fusion gene and has nucleotide sequences as shown in SEQ ID NO: 7-8; the probes correspond to the three groups of primer pairs. The method not only realizes early diagnosis of the prostate cancer, but also realizes effective evaluation of metastasis and deterioration risks of the prostate cancer of a diagnosed patient, and is suitable for large-scale population screening and general survey.

Description

technical field [0001] The invention relates to the field of molecular biology technology, and more specifically relates to a primer-probe combination, a kit and an application thereof for early diagnosis of prostate cancer, metastasis early warning and prognosis assessment. Background technique [0002] Prostate cancer refers to epithelial malignant tumors that occur in the prostate. "Pathology and Genetics of Tumors of the Urinary System and Male Reproductive Organs" published by WHO divides the pathological types of prostate cancer into adenocarcinoma (acinar adenocarcinoma), ductal adenocarcinoma, urothelial carcinoma, squamous cell carcinoma, and adenosquamous carcinoma . Among them, prostate adenocarcinoma accounts for more than 95%. Therefore, what we usually call prostate cancer refers to prostate adenocarcinoma. [0003] In 2020, a total of 1.41 million people will be diagnosed with prostate cancer and 380,000 will die, ranking fourth in the incidence rate of canc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/118C12Q2600/158C12Q2600/166C12Q2531/113C12Q2547/101C12Q2521/107C12Q2563/107C12Q2545/101
Inventor 何熲何小明张辉包海波巩许婷韦鑫邱伟
Owner SHANGHAI CONLIGHT MEDICAL LTD
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