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Kit and method for detecting BK virus in urine

A kit and virus technology, which is applied in the field of kits for detecting urine BK virus, can solve the problems of low detection accuracy, slow speed, and long BK virus detection cycle, and achieve short cycle, high accuracy, and specific detection sex high effect

Pending Publication Date: 2020-08-18
南京实践医学检验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Most of the existing methods for detecting BK virus have a long detection cycle, slow speed, and low detection accuracy, so we urgently need products that can quickly, effectively and accurately detect BK virus for the detection of BK virus load

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0030] A kit for detecting urine BK virus, which includes 2×Taq Mix, BK virus amplification primers, ddH 2 O; the 2 × Taq Mix includes TaqDNA polymerase, dNTPs, standard Taq enzyme reaction buffer, enzyme stabilizer and bromophenol blue dye,

[0031] The BK virus amplification primers are:

[0032] F: GGGCAGCGCACGATGTTTT;

[0033] R: AGCTGCCCCTGGACACTCT.

[0034] A method for detecting urine BK virus, comprising the following steps in sequence:

[0035] Step 1: Obtaining the sample to be tested: extract the DNA of BK virus from the urine to be tested to obtain the sample to be tested, and the specific steps for extracting DNA from the urine are as follows:

[0036] Lysis: Add 200 μL lysate and 20 μL biog compound digestion solution to the sample, mix well, and incubate at 56°C for 12 minutes;

[0037] Binding: Cool to room temperature, add 300 μL isopropanol, mix thoroughly, then add 20 μL magnetic bead complex solution, oscillate and mix for 3 minutes, rest for 2 minutes,...

Embodiment 2

[0047] A kit for detecting urine BK virus, which includes 2×Taq Mix, BK virus amplification primers, ddH 2O; the 2 × Taq Mix includes TaqDNA polymerase, dNTPs, standard Taq enzyme reaction buffer, enzyme stabilizer and bromophenol blue dye,

[0048] The BK virus amplification primers are:

[0049] F: GGGCAGCGCACGATGTTTT;

[0050] R: AGCTGCCCCTGGACACTCT.

[0051] A method for detecting urine BK virus, comprising the following steps in sequence:

[0052] Step 1: Obtaining the sample to be tested: extract the DNA of BK virus from the urine to be tested to obtain the sample to be tested, and the specific steps for extracting DNA from the urine are as follows:

[0053] Lysis: Add 200 μL lysate and 20 μL biog compound digestion solution to the sample, mix well, and incubate at 56°C for 12 minutes;

[0054] Binding: Cool to room temperature, add 300 μL isopropanol, mix thoroughly, then add 20 μL magnetic bead complex solution, oscillate and mix for 3 minutes, rest for 2 minutes, ...

Embodiment 3

[0064] A kit for detecting urine BK virus, which includes 2×Taq Mix, BK virus amplification primers, ddH 2 O; the 2 × Taq Mix includes TaqDNA polymerase, dNTPs, standard Taq enzyme reaction buffer, enzyme stabilizer and bromophenol blue dye,

[0065] The BK virus amplification primers are:

[0066] F: GGGCAGCGCACGATGTTTT;

[0067] R: AGCTGCCCCTGGACACTCT.

[0068] A method for detecting urine BK virus, comprising the following steps in sequence:

[0069] Step 1: Obtaining the sample to be tested: extract the DNA of BK virus from the urine to be tested to obtain the sample to be tested, and the specific steps for extracting DNA from the urine are as follows:

[0070] Lysis: Add 200 μL lysate and 20 μL biog compound digestion solution to the sample, mix well, and incubate at 56°C for 12 minutes;

[0071] Binding: Cool to room temperature, add 300 μL isopropanol, mix thoroughly, then add 20 μL magnetic bead complex solution, oscillate and mix for 3 minutes, rest for 2 minutes,...

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Abstract

The invention discloses a kit and a method for detecting a BK virus in urine. The kit comprises a 2*Taq Mix, a BK virus amplification primer and ddH2O, wherein the BK virus amplification primer is asfollows: F: GGGCAGCGCACGATGTTTT; R: AGCTGCCCCTGGACACTCT. The detection method comprises the following steps in sequence: 1: obtaining a sample to be detected; 2, preparing a PCR reaction solution; 3,carrying out PCR amplification; 4, recovering a target strip; 5, purifying the DNA recovered from the sample, and then connecting a vector for sequencing; and 6, comparing the measured sequence with the sequence of the BK virus, indicating that the BK virus exists in the sample if the sequences are consistent, and indicating that the BK virus does not exist in the sample if the sequences are inconsistent or no strip with the same size as the positive control exists in a sample tube. The kit and the method for detecting the BK virus in urine can rapidly detect whether the urine contains the BKvirus or not, are high in accuracy and short in period, and adopt a specific BK primer, so that the detection specificity is high.

Description

technical field [0001] The invention belongs to the technical field of biological virus detection, in particular to a kit and method for detecting urine BK virus. Background technique [0002] Human polyomavirus type BK (BK virus) is a non-enveloped virus with a circular double-stranded DNA genome of approximately 5300 bp. BK virus was first recognized as a member of the polyomavirus family in 1971 after isolation from the urine of kidney transplant recipients. Subsequent studies have shown that the seropositive rate of adults worldwide is over 80%. Typically, primary infection with BK virus occurs during childhood through the respiratory tract, with subsequent viral dormancy in the genitourinary tract. Asymptomatic reactivation and intermittent shedding of virus in urine occurs naturally in immunocompetent persons, but in those with altered cellular immunity such as pregnant women, cancer patients undergoing chemotherapy, HIV-1-infected individuals, and renal or other Th...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6869C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6869
Inventor 张鹏唐春花邢宽何志健谢珍倪海钰周权何贵伦安雪茹李平江晓琴
Owner 南京实践医学检验有限公司
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