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Novel method used for quantification of baculovirus using high performance liquid chromatographic instrument

A high-performance liquid chromatography, baculovirus technology, applied in instruments, measuring devices, scientific instruments, etc., can solve problems such as error in detection results, and achieve the effects of reducing experimental costs, improving experimental efficacy, and shortening inspection cycles.

Inactive Publication Date: 2014-04-23
TECON BIOLOGY CO LTD
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  • Abstract
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Problems solved by technology

The detection period of this method is 8 to 9 days, and the determination of the poison price is affected by personnel and equipment, resulting in differences, resulting in errors in the detection results

Method used

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  • Novel method used for quantification of baculovirus using high performance liquid chromatographic instrument
  • Novel method used for quantification of baculovirus using high performance liquid chromatographic instrument
  • Novel method used for quantification of baculovirus using high performance liquid chromatographic instrument

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Embodiment Construction

[0036] The invention will be further described below in conjunction with examples.

[0037] Quantification of insect baculovirus by indirect immunofluorescence (the recombinant baculovirus of classical swine fever virus E2 is also a kind of insect baculovirus, which is described as an example);

[0038] Among them: dilution of recombinant baculovirus of classical swine fever virus E2

[0039] Take out 6 different batches of CSFV E2 recombinant baculovirus from -70 ℃ refrigerator and carry out serial 10-fold gradient dilution respectively with SF900II serum-free insect cell culture medium (10 -1 ~10 -11 ), that is, take 100 μl of classical swine fever virus E2 recombinant baculovirus and add it to 900 μl of SF900II serum-free insect cell culture medium, so dilute, each dilution takes 100 μl of diluted classical swine fever E2 recombinant baculovirus solution (hereinafter referred to as baculovirus solution ) to a 96-well plate (eight replicates for each dilution).

[0040] W...

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Abstract

The invention provides a novel method used for quantification of baculovirus using a high performance liquid chromatographic instrument. The novel method comprises following steps: dilution of baculovirus; immobilization of baculovirus; permeabilization of baculovirus; staining of baculovirus; construction of a standard curve and quantification of samples to be tested using the high performance liquid chromatographic instrument. In addition, dilution of baculovirus; immobilization of baculovirus; permeabilization of baculovirus; staining of baculovirus; determination of maximum absorption peak of samples using the high performance liquid chromatographic instrument; and calculating of viral titer based on the standard curve are performed at the same time. Compared with traditional methods, the novel method is capable of shortening baculovirus quantification time greatly, increasing sensitivity of virus quantification, reducing experiment cost, and increasing experiment efficiency.

Description

technical field [0001] The invention relates to the quantitative analysis of the insect baculovirus, and the adopted new method shortens the quantitative time and sensitivity of the baculovirus, reduces the experiment cost and improves the experiment effect. Background technique [0002] The conventional quantitative method of insect baculovirus is to use cell culture medium to serially dilute the baculovirus by 10 times. The plate was cultured in a constant temperature incubator at 27°C for about 7 days, and the virus titer was determined by indirect immunofluorescence. The detection period of this method is 8-9 days, and the determination of the poison price is affected by personnel and equipment, resulting in differences, resulting in errors in the detection results. [0003] The quantitative SOP of the insect baculovirus disclosed by literature search: the rod is serially diluted 10 times with the insect cell culture medium (10 -1 ~10 -11 ), add 100 μl of each dilutio...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 李俊辉赵毅
Owner TECON BIOLOGY CO LTD
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