CN102191307A - Method for enriching Cryptosporidium parvum oocysts and Giardia Lamblia sporocysts in water - Google Patents
Method for enriching Cryptosporidium parvum oocysts and Giardia Lamblia sporocysts in water Download PDFInfo
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- CN102191307A CN102191307A CN2010101267454A CN201010126745A CN102191307A CN 102191307 A CN102191307 A CN 102191307A CN 2010101267454 A CN2010101267454 A CN 2010101267454A CN 201010126745 A CN201010126745 A CN 201010126745A CN 102191307 A CN102191307 A CN 102191307A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The invention relates to a method for enriching Cryptosporidium parvum oocysts and Giardia Lamblia sporocysts in water. The method comprises the following steps: adding an inorganic metal salt coagulant into a bulk water sample, stirring, coagulating, precipitating, collecting precipitate alumen ustum in a small-volume container, adding acidic solution into the small-volume container to dissolve the alumen ustum, centrifuging, removing supernate and thus, enriching the Cryptosporidium parvum oocysts and Giardia Lamblia sporocysts in water. In the invention, the problems of low Cryptosporidium parvum oocyst and Giardia Lamblia sporocyst enrichment rate, instability, high operation requirement and expensive filtering equipment and consumable items of the conventional method for enriching Cryptosporidium parvum oocysts and Giardia Lamblia sporocysts. The method does not need expensive apparatus and equipment and medicines and greatly reduce capital investment. The method is simple in operation, easy to master, time-saving and labor-saving, reduces cost of enrichment of Cryptosporidium parvum oocysts and Giardia Lamblia sporocysts in water, improves enrichment efficiency and is a novel method which clears the preliminary work for subsequent steps of immunomagnetic separation, immunofluorescence label microscope counting, polymerase chain reaction (PCR) qualitative detection, nano probe gene chip detection and Real-time PCR quantitative detection and the like.
Description
Technical field
The present invention relates to the method in a kind of detection technique field, the method for Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst belongs to the water quality safety technical field in especially a kind of coagulation enrichment water.
Background technology
Cryptosporidium parvum (Cryptosporidium parvum) and Giardia lamblia (Giardia lamblia) are considered to most important two kinds of water at present and pass enteropahtogenic microganism.Cryptosporidium is present in the water with the form of egg capsule (Oocyst), and median infective dose is 132 egg capsules alive, and pathogenic amount is generally 10 egg capsules alive; And giardia lamblia stiles is present in the water with the form of sporangiocyst (Cyst), and pathogenic dosage is generally 10～25 sporangiocysts alive.Drunk the water that contains " two worms " (cryptosporidium parvum and Giardia lamblia) and can cause digestive tract infection, with symptoms such as stomachache, diarrhoea, heatings.Jie's water of " two worms " is propagated and can be caused serious public safety incident.At present in the world to still not having effective methods of treatment because of " two worms " infects the intestinal tract disease that causes." two worms " constitutes a serious threat to drinking water safety, caused the concern of the countries in the world that comprise China, in " the drinking water sanitary standard GB 5749-2006 " of the new promulgation of China in 2006, stipulated that clearly the level of " two worms " in the tap water should be in per 10 premium on currency less than 1 egg capsule or sporangiocyst.
American National Environmental Protection Administration (USEPA) formulates and 1623 methods of issue are present the most frequently used in the world " two worms " examination criteria methods, this method is at first by two worms in the filteration trap water, use washings will be trapped in " two worms " washing on filter capsule surface subsequently, adopt methods such as immune magnetic separation, DAPI dyeing and differential interference microscope observation counting to carry out quantitative assay subsequently.This method causes the loss of " two worms " in the process of wash-out filter capsule, make that the accumulation rate of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst is low, studies show that the rate of recovery of Cryptosporidium egg capsule and giardia lamblia sporangiocyst often has only 20%～30% lower level.This method exists operational requirement height, filter plant and consumptive material (as special use filter capsule etc.) and costs an arm and a leg, and two worm accumulation rates are low and problem such as instability.
Therefore, be necessary to develop a kind of simple to operate, time saving and energy saving, can reduce the method for the cost of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in the enrichment water, improve the accumulation rate of " two worms ", for steps such as follow-up immune magnetic separation, immunofluorescence label microscopic counting, PCR qualitative detection, the detection of nano-probe gene chip and Real-time PCR detection by quantitative provide good assurance.
Summary of the invention
The purpose of this invention is to provide a kind of easy, enriching method of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in the water fast and efficiently.
The step of present method is as follows:
(1) in water sample, adds 10～100mg/l inorganic metal salt coagulant;
(2) stir postprecipitation, abandoning supernatant is collected sedimentary alumen ustum;
(3) in alumen ustum, add acid solution and make the alumen ustum dissolving;
(4) solution is after centrifugal, and abandoning supernatant gets final product.
Described inorganic metal salt coagulant is: aluminum chloride, Tai-Ace S 150, iron(ic) chloride, ferric sulfate, polymerize aluminum chloride, poly-ferric chloride and aluminium iron polychloride.
Described acid solution is: hydrochloric acid, sulfuric acid.
Metallic salt coagulating agent of the present invention abundant hydrolysis in water forms the solvability polymkeric substance, and then the electrostatic double layer of the organic polymer ionogen (as humic acid and fulvic acid) of electronegative clay colloid, electronegative functional group in compression and the water body, make the water-borne glue body take off steady polymerization; " two worms " particle is slightly electronegative, can be caught by the hydroxide precipitate net, together precipitates with flco.This process is transferred to " two worms " in the water in the less flco throw out of volume, has realized the preliminary enrichment of " two worms ".
Then add acid solution in the alumen ustum solution that contains two worms after collecting, feasible alumen ustum generation acid-base neutralisation reaction based on metal hydroxides, thus make the alumen ustum dissolving, " two worms " is released in the solution of small volume once more.Again with whizzer to the solution centrifugal after concentrating through vast scale, thereby reach the purpose of efficiently concentrating " two worms ".
Advantage of the present invention and positively effect: the present invention need not expensive plant and instrument, medicine, has greatly reduced fund input.This method is simple to operate, time saving and energy saving, reduced the cost of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in the enrichment water, and improved accumulation rate, be a kind of novel method that previous work is provided for steps such as follow-up immune magnetic separation, immunofluorescence label microscopic counting, PCR qualitative detection, the detection of nano-probe gene chip and Real-time PCR detection by quantitative.
Embodiment:
Below in conjunction with specific embodiment technical scheme of the present invention is described in further detail.
Embodiment 1
With the tap water is water sample, uses polymerize aluminum chloride (PAC) and aluminium iron polychloride (PAFC) to be coagulating agent, and Cryptosporidium egg capsule and the giardia lamblia stiles sporangiocyst that adds in the water sample carried out the coagulation enrichment.The water sample amount is 1L from the beginning, adds Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst respectively, makes to contain about 1000 of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in the water, and water sample pH value is 9.0; The PAC dosage is that 30mg/L, PAFC dosage are 20mg/L, and agitation condition is to stir 180r/min2min fast, stirs 40r/min 15min more at a slow speed, and reaction is left standstill 30min after finishing.
Abandoning supernatant is collected sedimentary alumen ustum subsequently, adds the hydrochloric acid of 1mL 1.0mol/L in collected alumen ustum, stirs to make the alumen ustum dissolving.With this solution under 10000 rev/mins of conditions centrifugal 10 minutes, subsequently with the supernatant liquid sucking-off, keep centrifuge tube bottom settlings thing, promptly realized the enrichment of " two worms " in the water.
Table 1 has provided the accumulation rate of the present invention to Cryptosporidium egg capsule in the tap water and giardia lamblia stiles sporangiocyst.
Table 1 the present invention is to the accumulation rate of Cryptosporidium egg capsule in the tap water and giardia lamblia stiles sporangiocyst
Embodiment 2
With apricot woods gulf, Xiamen City reservoir water is sample, use Tai-Ace S 150, ferric sulfate, iron(ic) chloride, PAC, PAFC to be coagulating agent, amount is the alternative indicator (fluorescent microsphere of two worms of 10000/L in the water sample to adding, be divided into Φ 6.0 μ m, be used to simulate Cryptosporidium egg capsule and Φ 10.0 μ m, be used to simulate Lan Shi giardia lamblia sporangiocyst, purchase Polysciences company to the U.S.) carry out the coagulation enrichment.Enriching method is with embodiment 1.
Table 2 has provided in the example 2 the present invention to the accumulation rate of Cryptosporidium microballoon in the surface water and giardia lamblia stiles microballoon.
Table 2 different coagulants is to the Cryptosporidium microballoon that adds in the surface water and the accumulation rate of giardia lamblia stiles microballoon
By above embodiment as seen, the present invention is 73.8-92.9% to the accumulation rate of Cryptosporidium egg capsule in the tap water, the accumulation rate of giardia lamblia stiles sporangiocyst is 73.8-78.6%, and Cryptosporidium microballoon and giardia lamblia stiles microballoon accumulation rate can reach 99.99% in the surface water.The invention solves " two worms " of original EPA1623 method accumulation rate be low and unstable, operational requirement is high, filter plant and the expensive problem of consumptive material.And the present invention does not need expensive plant and instrument, medicine, greatly reduce fund input, easy grasp simple to operate, time saving and energy saving, can reduce the cost of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in the enrichment water, and the raising accumulation rate, be a kind of novel method that previous work is provided for steps such as follow-up immune magnetic separation, immunofluorescence label microscopic counting, PCR qualitative detection, the detection of nano-probe gene chip and Real-time PCR detection by quantitative.
Claims (3)
1. the enriching method of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in the water is characterized in that the step of method is as follows:
(1) in water sample, adds 10～100mg/l inorganic metal salt coagulant;
(2) stir postprecipitation, abandoning supernatant is collected sedimentary alumen ustum;
(3) in alumen ustum, add acid solution and make the alumen ustum dissolving;
(4) solution is after centrifugal, and abandoning supernatant gets final product.
2. the enriching method of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in a kind of water according to claim 1 is characterized in that described inorganic metal salt coagulant is: aluminum chloride, Tai-Ace S 150, iron(ic) chloride, ferric sulfate, polymerize aluminum chloride, poly-ferric chloride or aluminium iron polychloride.
3. the enriching method of Cryptosporidium egg capsule and giardia lamblia stiles sporangiocyst in a kind of water according to claim 1 is characterized in that described acid solution is: hydrochloric acid or sulfuric acid.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104020012A (en) * | 2014-06-25 | 2014-09-03 | 山东绿洁环境检测有限公司 | Sampling filtering bag for cryptospsridium and giardia detection in water |
| CN104726337A (en) * | 2013-12-20 | 2015-06-24 | 中国科学院生态环境研究中心 | Method for enriching and purifying cryptosporidium and giardia in alga-containing water, and method for determining content of cryptosporidium and giardia |
| CN107569889A (en) * | 2017-10-12 | 2018-01-12 | 西安思坦科技有限公司 | On Cryptosporidium and the filtering recovering device and its system of giardia lamblia stiles |
| CN109526904A (en) * | 2018-12-06 | 2019-03-29 | 华东师范大学 | A kind of water body control mosquito device and control mosquito method luring mosquito and coagulation mosquito eradication based on multistage |
| CN109959551A (en) * | 2019-02-28 | 2019-07-02 | 华东理工大学 | Using the method and application of parasitic ovum (spore) capsule in flocculation technique enrichment water sample |
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| CN1908621A (en) * | 2006-08-17 | 2007-02-07 | 上海交通大学 | Giardia sporangiocyst in water and Cryptosporidium hominis egg capsule checking method |
| CN101430281A (en) * | 2008-12-17 | 2009-05-13 | 哈尔滨工业大学 | Active detection method for concealed spore egg vesicle and Giardia sporocyst in drinking water |
| CN101665820A (en) * | 2009-09-15 | 2010-03-10 | 上海交通大学 | Filtration and concentration method for improving detection recovery rate of cryptosporidium and giardia |
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2010
- 2010-03-11 CN CN2010101267454A patent/CN102191307A/en active Pending
Patent Citations (3)
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| CN1908621A (en) * | 2006-08-17 | 2007-02-07 | 上海交通大学 | Giardia sporangiocyst in water and Cryptosporidium hominis egg capsule checking method |
| CN101430281A (en) * | 2008-12-17 | 2009-05-13 | 哈尔滨工业大学 | Active detection method for concealed spore egg vesicle and Giardia sporocyst in drinking water |
| CN101665820A (en) * | 2009-09-15 | 2010-03-10 | 上海交通大学 | Filtration and concentration method for improving detection recovery rate of cryptosporidium and giardia |
Non-Patent Citations (3)
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| TETSUJI OKUDA 等: "Enhancement of cryptosporidium oocyst removal by coagulation and sedimentation with poly-silicate iron (PSI)", 《JOURNAL OF CHEMICAL ENGINEERING OF JAPAN》 * |
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| 蔡炯 等: "生活饮用水中贾第鞭毛虫和隐孢子虫定量检测的质量控制", 《现代预防医学》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104726337A (en) * | 2013-12-20 | 2015-06-24 | 中国科学院生态环境研究中心 | Method for enriching and purifying cryptosporidium and giardia in alga-containing water, and method for determining content of cryptosporidium and giardia |
| CN104020012A (en) * | 2014-06-25 | 2014-09-03 | 山东绿洁环境检测有限公司 | Sampling filtering bag for cryptospsridium and giardia detection in water |
| CN107569889A (en) * | 2017-10-12 | 2018-01-12 | 西安思坦科技有限公司 | On Cryptosporidium and the filtering recovering device and its system of giardia lamblia stiles |
| CN109526904A (en) * | 2018-12-06 | 2019-03-29 | 华东师范大学 | A kind of water body control mosquito device and control mosquito method luring mosquito and coagulation mosquito eradication based on multistage |
| CN109959551A (en) * | 2019-02-28 | 2019-07-02 | 华东理工大学 | Using the method and application of parasitic ovum (spore) capsule in flocculation technique enrichment water sample |
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Application publication date: 20110921 |