Method for enriching water body hepatitis A viruses based on mixed cellulose ester membrane

A technology of mixing cellulose and hepatitis A virus, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/testing, etc., can solve the problems of incomparability and great difference in the results of enrichment methods, and achieve simple operation Easy to operate, good physical retention effect and high binding rate

Inactive Publication Date: 2014-05-14
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] Due to the different physical and chemical properties of materials and materials, adsorption-elution conditions, and operating procedures in various enrichment methods, the results of different enrichment methods are also very different, and countries are not comparable.

Method used

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  • Method for enriching water body hepatitis A viruses based on mixed cellulose ester membrane
  • Method for enriching water body hepatitis A viruses based on mixed cellulose ester membrane
  • Method for enriching water body hepatitis A viruses based on mixed cellulose ester membrane

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Experimental program
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Effect test

Embodiment 1

[0038] The establishment of the method for the enrichment and detection of hepatitis A virus in PBS water samples, the specific steps are as follows:

[0039] Before enriching the virus in the water body, the virus enrichment container etc. should be sterilized (sterilized by UV for 24 hours or sterilized by high temperature and high pressure).

[0040] 1. Enrichment of Hepatitis A virus in PBS water sample by using mixed cellulose ester membrane

[0041] Enrichment device ( figure 1 ): Vacuum pump 1 (purchased from Tianjin Autosines Instrument Co., Ltd.), connecting tube 2, filter 3 (Nalgene) and filter membrane 4. The vacuum pump 1 communicates with the filter 3 through the connecting pipe 2, and the filter membrane 4 is fixed in the filter 3. The device is easy to operate, light and suitable for laboratory operation.

[0042] Dilute 1ml of hepatitis A virus (i.e. freeze-dried live attenuated hepatitis A vaccine, purchased from Institute of Medical Biology, Chinese Academ...

Embodiment 2

[0061] The enrichment and detection of Hepatitis A virus in water samples from Donghu Lake, the specific steps are as follows:

[0062] Before sampling and enriching the virus in the water body, the water sample collection container and the virus enrichment container should be sterilized (sterilized by UV for 24 hours or sterilized by high temperature and high pressure).

[0063] 1. East Lake water sample pretreatment

[0064] Water samples from East Lake were collected, and 1ml of hepatitis A virus (i.e. freeze-dried live attenuated hepatitis A vaccine, purchased from the Institute of Medical Biology, Chinese Academy of Medical Sciences) and PBS components (137mmol / L NaCl, 2.7mmol / L NaCl, 2.7mmol / L L KCl, 10mmol / L Na 2 HPO 4 and 2mmol / L KH 2 PO 4 ), use HCl to adjust the pH of the solution to 7.2-7.4; set up a negative control East Lake water sample, add PBS components and use HCl to adjust the pH of the solution to 7.2-7.4, without adding hepatitis A virus; the positive ...

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Abstract

The invention provides a method for enriching water body hepatitis A viruses based on a mixed cellulose ester membrane. The method specifically comprises the following steps: pre-treating a water sample; filtering the water sample by using a glass fiber membrane; enriching the hepatitis A viruses in the water sample by using the mixed cellulose ester membrane; extracting RNA (Ribonucleic Acid) of the viruses and carrying out reverse transcription; manufacturing a standard curve; detecting the hepatitis A viruses by using fluorescence quantitative PCR (Polymerase Chain Reaction); calculating the enriching efficiency of the hepatitis A viruses. The method provided by the invention is reasonable in concept; the device is simple to operate, is light and is suitable for laboratory operation and wild work; conventional secondary concentration of adding an eluting agent to elute the viruses and adding an organic or inorganic flocculent precipitation agent to precipitate the viruses is saved and the volume can be concentrated for about 250 times; particularly, the enriching efficiency is high and the enriching efficiency in a PBS (Phosphate Buffered Saline) water sample averagely reaches (92.62+/-5.17)%; the enriching efficiency in an East Lake water sample averagely reaches (79.45+/-12.10)%. The method has very important meanings on efficient and accurate detection and monitoring on the hepatitis A viruses in the water bodies of various environments.

Description

technical field [0001] The invention belongs to the technical field of biological analysis testing and environmental monitoring, and specifically relates to a method for enriching hepatitis A virus in water based on a mixed cellulose ester film and its application. Fluorescent quantitative PCR was used to detect its copy number, so as to improve the sensitivity of detecting virus pollution in water. Background technique [0002] In recent years, as the environmental pollution has become more and more serious, pathogenic diseases caused by water transmission have become one of the problems that people pay close attention to worldwide. At present, many countries, including China, use fecal pollution indicator bacteria, such as fecal coliforms, total coliforms, and enterococci, to evaluate the pollution status of water bodies. my country's current "Drinking Water Standards" (GB5742-2006) and wastewater reuse documents "Reclaimed Water Quality Standards" (SL368-2006) only inclu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/02C12Q1/70C12Q1/68C12R1/93
Inventor 陈全姣乔煜婷隋志伟高志敏赵丽华
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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