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Urine detection kit for prostate cancer and application thereof

A detection kit and technology for prostate cancer, applied in the detection field, can solve the problems of complex quality control and result interpretation, easy pollution of the detection environment, false positive detection results, etc., to avoid false positive phenomena, and to achieve simple and accurate quality control methods. Good results

Active Publication Date: 2015-05-20
JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is cumbersome, time-consuming, complex in quality control and result interpretation, and has a high retest rate of 15.2%. It is incompatible with clinical routine PCR instruments, and additional equipment needs to be purchased
More importantly, the detection process needs to be repeatedly opened to add reagents, which is easy to pollute the detection environment, resulting in false positives in subsequent detection results

Method used

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  • Urine detection kit for prostate cancer and application thereof
  • Urine detection kit for prostate cancer and application thereof
  • Urine detection kit for prostate cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 kit detection reagent, detection condition lower limit detection ability

[0033] 1. Prostate cancer urine detection kit: including sample transportation preservation solution, magnetic bead method extraction reagent, one-step RT-PCR amplification reagent, standard product, positive quality control product, and negative quality control product.

[0034] Sample transport preservation solution: the composition is 1% lithium dodecyl sulfate by mass fraction, 10mM NaH 2 PO 4 , 10mM Na 2 HPO 4 , 0.5mM ethylenediaminetetraacetic acid (EDTA), 0.5mM ethylene glycol diethyl ether diamine tetraacetic acid (EGTA), pH6.7;

[0035] Magnetic bead method extraction reagent:

[0036] (1) Capture solution: 100mM HEPES, 0.5M LiCl, 100mM LiOH, 50mM EDTA and 50μg / mL superparamagnetic covalently bound (dT)14 magnetic beads (0.7~1.05μMparticles, Sera-Mag TM ), pH6.4;

[0037] (2) Lotion: 5mM HEPES, 100mM NaCl, 4mM NaOH, 0.5mM EDTA, 0.1% (v / v) ethanol, 0.01% (w / v) methyl p-...

Embodiment 2

[0046] Embodiment 2 Kit detection reagent, detection condition upper limit detection ability

[0047] 1. Prostate cancer urine detection kit: including sample transportation preservation solution, magnetic bead method extraction reagent, one-step RT-PCR amplification reagent, standard product, positive quality control product, and negative quality control product.

[0048] Sample transport preservation solution: the composition is 5% lithium dodecyl sulfate, 20mM NaH 2 PO 4 , 20mM Na 2 HPO 4 , 2mM ethylenediaminetetraacetic acid (EDTA), 2mM ethylene glycol diethyl ether diamine tetraacetic acid (EGTA), pH6.7;

[0049] Magnetic bead method extraction reagent:

[0050] (1) Capture solution: 400mM HEPES, 3M LiCl, 500mM LiOH, 150mM EDTA and 400μg / mL superparamagnetic covalently bound (dT)14 magnetic beads (0.7~1.05μMparticles, Sera-Mag TM ), pH6.4;

[0051] (2) Lotion: 20mM HEPES, 200mM NaCl, 10mM NaOH, 2mM EDTA, 0.5% (v / v) ethanol, 0.02% (w / v) methyl p-hydroxybenzoate, 0.05...

Embodiment 3

[0061] 1. Prostate cancer urine detection kit: including sample transportation preservation solution, magnetic bead method extraction reagent, one-step RT-PCR amplification reagent, standard product, positive quality control product, and negative quality control product.

[0062] Sample transport preservation solution: the composition is 3% lithium dodecyl sulfate, 15mM NaH 2 PO 4 , 15mM Na 2 HPO 4 , 1mM ethylenediaminetetraacetic acid (EDTA), 1mM ethylene glycol diethyl ether diamine tetraacetic acid (EGTA), pH6.7;

[0063] Magnetic bead method extraction reagent:

[0064] (1) Capture solution: 250mM HEPES, 1.88M LiCl, 310mM LiOH, 100mM EDTA and 250μg / mL superparamagnetic covalently bound (dT)14 magnetic beads (0.7-1.05μM particles, Sera-Mag TM ), pH6.4;

[0065] (2) Lotion: 10mM HEPES, 150mM NaCl, 6.5mM NaOH, 1mM EDTA, 0.3% (v / v) ethanol, 0.02% (w / v) methyl p-hydroxybenzoate, 0.01% (w / v) nitric acid Propyl Parkinate and 0.1% (w / v) Sodium Lauryl Sulfate, pH 7.5.

[006...

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Abstract

The invention discloses a urine detection kit for prostate cancer. The kit comprises a one-step method RT-PCR (Reverse Transcription-Polymerase Chain Reaction) amplification reagent which comprises specific primers for detecting PCA (Prostate Cancer Antigen) 3 and PSA (Prostate Specific Antigen) and a probe. The primers for detecting the PCA3 are as shown in SEQ ID NO.1 and SEQ ID NO.2, and the probe is as shown in SEQ ID NO.5. The primers for detecting the PSA are as shown in SEQ ID NO.3 and SEQ ID NO.4, and the probe is as shown in SEQ ID NO.6. The kit has the advantages of simple and convenient operational process, short consumed time, simplicity in result judgment, good repeatability and low recheck rate, and after a template is added, the whole course of the PCR reaction is carried out in a closed tube, so that a false positive phenomenon caused by pollution is avoided.

Description

technical field [0001] The invention belongs to the detection field, in particular to a urine detection kit for prostate cancer, and also relates to the application of the kit. Background technique [0002] Prostate cancer is a malignant tumor that occurs in the prostate tissue of men and is the result of abnormal and disordered growth of prostate acinar cells. Prostate cancer incidence varies markedly by geographic and ethnic origin. In developed countries and regions such as Europe and the United States, it is the most common malignant tumor in men, and its mortality rate ranks second among various cancers, second only to lung cancer; in Asia, its incidence rate is lower than that of Western countries, but it has increased rapidly in recent years Upward trend. Prostate cancer usually occurs after the age of 50, and 95% occurs in elderly men over the age of 60, and the incidence rate increases with age. There are 70,000 to 80,000 new prostate cancer patients in my countr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2531/113
Inventor 王弢渠香云何林富高鹏
Owner JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD
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