Method for enriching and purifying two insects in water and method for measuring content of insects

A purification method and enrichment technology, which can be used in microorganism-based methods, biochemical equipment and methods, and the determination/inspection of microorganisms, which can solve the problems of high experimental cost, troublesome operation, and low recovery rate.

Active Publication Date: 2014-02-12
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under the condition of high turbidity in the source water, no matter whether the method of filter capsule / filter element or membrane filtration is used for concentration, multiple filter capsules / filter elements or filter membranes are often required, which is not only troublesome in operation, but also expensive in experiments; at the same time Due to excessive precipitation, it is not conducive to the further separation and purification of Cryptosporidium oocysts and Giardia cysts, resulting in a low recovery rate

Method used

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  • Method for enriching and purifying two insects in water and method for measuring content of insects
  • Method for enriching and purifying two insects in water and method for measuring content of insects
  • Method for enriching and purifying two insects in water and method for measuring content of insects

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1 Preparation of buffer solution

[0107] 1. Preparation of phosphate buffered saline solution (PBS solution):

[0108] 1) Prepare materials according to the following dosage (based on 1L water volume)

[0109] NaCl (analytical pure) 8.0g

[0110] Na 2 HPO 4 12H 2 O (analytical pure) 2.9g

[0111] KCl (analytical pure) 0.2g

[0112] K H 2 PO 4 (analytical pure) 0.2g

[0113] 2) Add the raw materials of the above ingredients into 1000mL of pure water, stir and dissolve evenly, and adjust the pH to 7.2-7.4 to obtain the product.

[0114] 2. Preparation of phosphate Tween buffer solution (PBST solution)

[0115] Add Tween 80 to the above-prepared PBS solution, stir and mix well to obtain, wherein 0.01-0.2 mL Tween 80 is added to every 100 mL of PBS solution.

[0116] In the embodiment of the present invention, 0.1 mL of Tween 80 was added to every 100 mL of PBS solution.

[0117] 3. Percoll-sucrose solution separation medium

[0118] Dissolve 45mL of Per...

Embodiment 2

[0123] 1) Prepare muddy water containing "two worms"

[0124] First, use kaolin to simulate turbidity to prepare turbid water, add 0.3485g kaolin to 10L deionized water, stir it with a magnetic stirrer to dissolve it completely, and obtain turbid water with a turbidity of 40NTU;

[0125] Then, add 500 Cryptosporidium oocysts and Giardia cysts "two worms" standard samples to the turbid water, so that every 1L of turbid water contains 50 Cryptosporidium oocysts and Giardia cysts ( 50 worms / L each), to obtain turbid water containing "two worms".

[0126] 2) Precipitation and enrichment of "two insects"

[0127] Add calcium chloride solution and sodium bicarbonate solution to 10L of turbid water samples containing "two worms", stir evenly with a magnetic stirrer, add 10mL of calcium chloride solution with a concentration of 1mol / L per 1L of turbid water, add 10 mL of sodium bicarbonate solution with a concentration of 1 mol / L;

[0128] Then add a sodium hydroxide solution with ...

Embodiment 3

[0150] Taking the source water of Huangpu River as a sample, its original turbidity is 56NTU.

[0151] 1) Precipitation and enrichment of "two insects"

[0152] Add calcium chloride solution and sodium bicarbonate solution to 10L of Huangpu River source water sample, stir evenly with a magnetic stirrer, add 20mL of calcium chloride solution with a concentration of 1mol / L to every 1L of turbid water, and add a concentration of 1mol / L 20 mL of sodium bicarbonate solution in L;

[0153] Then add a sodium hydroxide solution with a concentration of 10mol / L, adjust the pH of the turbid water sample to 10.00±0.05, and generate calcium carbonate precipitation under alkaline conditions, and the "two worms" sink to At the bottom, let it stand for 12 hours, remove the supernatant, and obtain a precipitate rich in "two worms".

[0154] 2) Acid dissociation treatment

[0155]Add sulfamic acid solution to the precipitate rich in "two worms" to dissolve the calcium carbonate precipitate t...

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Abstract

The invention relates to a method for enriching and purifying cryptosporidium and giardia in water and a method for measuring the content of the cryptosporidium and giardia. The method comprises the following steps: adding calcium chloride and sodium bicarbonate into a water sample, regulating the pH of the water sample, and settling and enriching the cryptosporidium and giardia along with the generated calcium carbonate under alkaline conditions; collecting the precipitate, adding a sulfamic acid solution, performing acid dissociation, and performing density gradient centrifugal treatment to obtain a purification solution of the cryptosporidium and giardia; and finally, adsorbing the cryptosporidium and giardia to a cellulose acetate membrane through filtering, performing fluorescence microscope detection after dyeing treatment, and counting. According to the method, a method for concentrating, separating and purifying the cryptosporidium and giardia in the water sample is improved, the interference of background impurities during microscopic examination is reduced, the detection method is high in recovery rate and accurate in detection result, and a quality control standard which is formulated in USA Environmental Protection Agency and is used for measuring the cryptosporidium and giardia in the water is met.

Description

technical field [0001] The invention relates to a method in the technical field of detection in water, in particular to a method for enriching and purifying Cryptosporidium oocysts and Giardia cysts in turbid water and a method for detecting their content in turbid water. Background technique [0002] Cryptosporidium and Giardia ("two worms") are two pathogenic protozoa that parasitize in humans and animals. In recent years, there have been many water-borne diseases caused by "two worms" in water abroad. The outbreak of the virus has attracted widespread attention. In my country's revised "Drinking Water Hygienic Standards" (GB5947-2006), the detection of Cryptosporidium and Giardia was added in the part of microbial indicators, and the limit value was clearly stipulated <1 / 10L. [0003] Through the search of the existing technology, it is found that the detection of "two insects" in water is accomplished through the main steps of water sample filtration and concentratio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/10C12Q1/06C12R1/90
Inventor 于建伟周美芝杨敏张冬青
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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