CN103045713B - Method for detecting trace amounts of microbes in a liquid sample containing multiple impurities - Google Patents
Method for detecting trace amounts of microbes in a liquid sample containing multiple impurities Download PDFInfo
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- CN103045713B CN103045713B CN201310018933.9A CN201310018933A CN103045713B CN 103045713 B CN103045713 B CN 103045713B CN 201310018933 A CN201310018933 A CN 201310018933A CN 103045713 B CN103045713 B CN 103045713B
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Abstract
The invention discloses a device and method for detecting trace amounts of microbes in a liquid sample containing multiple impurities. The method comprises the following steps: separating impurities and homogenizing the liquid with a membrane separation device and a homogenizer, carrying out vacuum filtration and enrichment with a microbial membrane filter, and finally, detecting after culturing. The device and method disclosed by the invention can be utilized to detect microbes in the massive liquid sample containing multiple solids and other large-granularity impurities as well as trace amounts of microbes, and have the advantages of high detection speed, favorable impurity removal accuracy and high efficiency.
Description
Technical field
The present invention relates to the detection method of a kind of microorganism, particularly the detection method of micro-microorganism in a kind of polymictic liquid sample.
Background technology
Other environment waters such as water of river and lake, process water, seawater are usually containing a large amount of solid impurity, a small amount of microorganism, in its clean level of evaluation or when needing certain microorganism of other aimed detection, is often difficult to be separated other impurity very well, and complex operation, easily produce error.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, the apparatus and method of micro-microorganism in a kind of many impurity liquid sample are provided.
For achieving the above object, the present invention takes following technical proposals to realize:
Micro-microorganism detection method in a kind of polymictic liquid sample, comprises the following steps:
A. get a certain amount of liquid sample to be detected, put into membrane separation unit, then membrane separation unit is placed in slap type homogenizer;
B., velocity of flapping and homogenizing time are set, are separated the solid impurity in testing sample, and make the liquid sample in membrane separation unit full and uniform;
C. to each parts of microbial film filtration unit, comprise stainless steel funnel and funnel web member carries out sterilizing, then connecting pipeline, closes each valve on stainless steel stent, opens vacuum pump, regulate vacuum tightness, with the tweezers after sterilization, sterilised membrane filter is placed on stainless steel filter membrane supporting pad, liquid sample after absorption separation homogeneous, to pour in stainless steel funnel to corresponding scale marks, open respective valves and carry out suction filtration;
D. after suction filtration completes, touch stainless steel funnel to side, then upwards take out stainless steel funnel, take off sterilised membrane filter with the antimagnetic type tweezer after sterilization, carry out corresponding detection;
E. the sterilised membrane filter after suction filtration enrichment is placed in the sterilized culture dish containing substratum, cultivates by inspecting standard, again detect after cultivation.
Further, to the liquid sample of small amount, quantitative sample and diluent are added in membrane separation unit simultaneously.
Preferably, to each parts of microbial film filtration unit, comprise stainless steel funnel and funnel web member carries out flame sterilization.
In polymictic liquid sample, a proofing unit for micro-microorganism, comprises membrane separation unit, slap type homogenizer, microbial film filtration unit.
Wherein, described membrane separation unit comprises the bag of two sides composition, bag four limit seals, top is can tear portions, bottom is bag main body, and inside is provided with filter membrane, and the base of described filter membrane and top are individually fixed in base and the top of bag, dual-side is individually fixed on the two sides of bag, and the width of filter membrane is less than the width of bag side.
Preferably, the dual-side of described filter membrane is individually fixed in the mid-way of bag two sides.
Wherein, described microbial film filtration unit comprises stainless steel stent, at least one stainless steel filter, stainless-steel vacuum table, vacuum filtration bottle, vacuum pump, wherein vacuum meter and stainless steel filter are installed on stainless steel stent by web member, vacuum filtration bottle is housed between stainless steel stent and vacuum pump, and stainless steel stent is connected by pipe connecting with vacuum filtration bottle, and described stainless steel filter comprises stainless steel base and is installed on the stainless steel funnel on stainless steel base.
Preferably, described stainless steel funnel is connected on stainless steel base by the mode of inlay card.
Preferably, described stainless steel base is provided with stainless steel filter membrane supporting pad.
The present invention has following technique effect: by adopting preceding method and device, isolate the solid in liquid sample and other large size impurity, then realize the full and uniform of sample, carry out suction filtration enrichment by microbial film filtration unit, finally detect through cultivation again, convenient detection contains many kinds of solids and other large granular impurities, and containing the microorganism detection in a large amount of liquid samples of a small amount of microorganism, detection speed is fast, despumation, accuracy is good, and efficiency is high;
Further, to the liquid sample of small amount, quantitative sample and diluent are added in membrane separation unit simultaneously, after dilution, follow-up suction filtration operation can being facilitated, simultaneously owing to having carried out homogeneous, without the need to worrying microbial profile heterogeneity, causing the problem of resultant error;
Further, before carrying out suction filtration, flame sterilization is carried out to microbial film filtration unit, rapidly and efficiently, can cool fast simultaneously, can drop into rapidly and reuse, save detection time, improve detection efficiency.
Accompanying drawing explanation
Fig. 1 is the schema of the detection method of micro-microorganism in a kind of polymictic liquid sample in the embodiment of the present invention.
Fig. 2 is the structural representation of microbial film filtration unit in the embodiment of the present invention.
Fig. 3 is the structural representation of membrane separation unit in the embodiment of the present invention.
Fig. 4 is the expansion schematic diagram of membrane separation unit in the embodiment of the present invention.
Embodiment
Below in conjunction with accompanying drawing, the present invention will be described in detail.
Fig. 1 is the schema of the detection method of micro-microorganism in a kind of polymictic liquid sample in the embodiment of the present invention.As shown in Figure 1, micro-microorganism detection method in this polymictic liquid sample, comprises the following steps:
A. pipette samples 11, gets a certain amount of liquid sample to be detected, puts into membrane separation unit, then membrane separation unit is placed in slap type homogenizer.
In the present embodiment, as more in testing liquid sample size, then directly can put into membrane separation unit, if when liquid sample amount is less, quantitative sample and diluent can be added in membrane separation unit, to reach the object of dilute sample simultaneously.
The present invention can select the multiple film tripping device and homogenizing bag etc. that are suitable for.Fig. 3 is the structural representation of the membrane separation unit adopted in the better embodiment of the present invention, Fig. 4 is the expansion schematic diagram of described membrane separation unit, composition graphs 3 and Fig. 4 can find out, described membrane separation unit comprises the bag of two sides composition, bag four limit seals, top is can tear portions 31, bottom is bag main body 32, inside is provided with filter membrane 33, the base of described filter membrane 33 and top are individually fixed in base and the top of bag, dual-side is individually fixed on the mid-way of the two sides of bag, and the width of filter membrane 33 is less than the width of bag side.
Membrane separation unit is divided into two spaces by described filter membrane 33, during application of sample, is added by testing sample in one of them space.
B. homogeneous is separated 12, arranges velocity of flapping and homogenizing time, is separated the solid impurity in testing sample, and makes the liquid sample in membrane separation unit full and uniform.
Difference per sample, can arrange suitable velocity of flapping and time, and bounce spacing etc.By the cooperation of slap type homogenizer, solid impurity in liquid sample in membrane separation unit and large-size particle are trapped within the space of the initial application of sample in filter membrane side, and the sample in the space of filter membrane opposite side removes clear solid impurity substantially, and microbial profile is even in sample, next step operation can be proceeded.
C. suction filtration enrichment 13, to each parts of microbial film filtration unit, comprise stainless steel funnel and funnel web member carries out sterilizing, then connecting pipeline, close each valve on stainless steel stent, open vacuum pump, regulate vacuum tightness, with the tweezers after sterilization, sterilised membrane filter is placed on stainless steel filter membrane supporting pad, draw and be separated liquid sample after homogeneous, to pour in stainless steel funnel to corresponding scale marks, open respective valves and carry out suction filtration.
In the present embodiment, multiple film filtration unit can be adopted, as long as the effect of aseptic enriched microorganism can be reached.Fig. 2 adopts microbial film filtration unit in good embodiment, comprise stainless steel stent 21, at least one stainless steel filter, stainless-steel vacuum table 22, vacuum filtration bottle 23, vacuum pump 24, wherein vacuum meter 22 and stainless steel filter 23 are installed on stainless steel stent 21, vacuum filtration bottle 25 is housed between stainless steel stent 21 and vacuum pump 24, stainless steel stent 21 is connected by pipe connecting 26 with vacuum filtration bottle 25, the stainless steel funnel 221 that described filter 22 comprises stainless steel base 222 and is installed on stainless steel base.
The filtration unit of aforementioned micro organism membrane filter appts adopts all steel material, the multiple sterilization methods such as moist heat sterilization, dry sterilization, flame sterilization can be selected, especially can select flame sterilization, rapidly and efficiently, and after sterilizing, speed of cooling is fast, can drop into rapidly and reuse.
In preferred embodiment, described stainless steel funnel is connected on stainless steel base by the mode of inlay card, concrete structure is: stainless steel base is provided with at least three stainless steel projections, when stainless steel funnel is subdued, projection is pressed into rear ejection, by funnel card jail, by the mode of this kind of inlay card, stainless steel funnel is fixed on stainless steel base, is convenient to picking and placeing of one-handed performance filter membrane.
Preferably, stainless steel base is provided with stainless steel filter membrane supporting pad, and this stainless steel filter membrane supporting pad is suppressed by stainless steel particle and formed, and supports evenly, ensure that the microorganism be trapped is uniformly distributed at filter membrane surface, be convenient to microorganism culturing subsequently, counting or grain size analysis etc.
D. detect the microorganism 14 after enrichment, after suction filtration completes, touch stainless steel funnel to side, then upwards take out stainless steel funnel, take off sterilised membrane filter with the antimagnetic type tweezer after sterilization, carry out corresponding detection.
Microorganism can reach finite concentration through suction filtration enrichment, according to actual needs, selects to carry out counting, detecting the operations such as analysis, also can proceed follow-up microorganism culturing.
E. follow-up cultivation detection 15, is placed in the sterilized culture dish containing substratum, cultivates, again detect after cultivation by inspecting standard by the sterilised membrane filter after suction filtration enrichment.
Although the present invention with preferred embodiment openly as above; but it is not for limiting the present invention; any those skilled in the art without departing from the spirit and scope of the present invention; the Method and Technology content of above-mentioned announcement can be utilized to make possible variation and amendment to technical solution of the present invention; therefore; every content not departing from technical solution of the present invention; according to technical spirit of the present invention to any simple modification made for any of the above embodiments, equivalent variations and modification, all belong to the protection of technical solution of the present invention.
Claims (6)
1. micro-microorganism detection method in a polymictic liquid sample, it is a kind of detection method for non-treatment and diagnostic purpose, it is characterized in that, comprise the following steps: get a certain amount of liquid sample to be detected, put into membrane separation unit, then membrane separation unit is placed in slap type homogenizer;
Velocity of flapping and homogenizing time are set, are separated the solid impurity in testing sample, and make the liquid sample in membrane separation unit full and uniform;
To each parts of the microbial film filtration unit of all steel, comprise stainless steel funnel and funnel web member carries out flame sterilization, then connecting pipeline, close each valve on stainless steel stent, open vacuum pump, regulate vacuum tightness, with the tweezers after sterilization, sterilised membrane filter is placed on stainless steel filter membrane supporting pad, liquid sample after absorption separation homogeneous, pour into corresponding scale marks in stainless steel funnel, open respective valves and carry out suction filtration, stainless steel filter membrane supporting pad is suppressed by stainless steel particle and is formed;
After suction filtration completes, touch stainless steel funnel to side, then upwards take out stainless steel funnel, take off sterilised membrane filter with the antimagnetic type tweezer after sterilization, carry out corresponding detection;
Sterilised membrane filter after suction filtration enrichment is placed in the sterilized culture dish containing substratum, cultivates by inspecting standard, again detect after cultivation.
2. detection method according to claim 1, is characterized in that, to the liquid sample of small amount, quantitative sample and diluent is added in membrane separation unit simultaneously.
3. method according to claim 1, is characterized in that, to each parts of microbial film filtration unit, comprises stainless steel funnel and funnel web member carries out flame sterilization.
4. the proofing unit of micro-microorganism in polymictic liquid sample, for the detection of non-treatment and diagnostic purpose, is characterized in that, comprise membrane separation unit, slap type homogenizer, and microbial film filtration unit; Described membrane separation unit comprises the bag of two sides composition, bag four limit seals, top is can tear portions, bottom is bag main body, inside is provided with filter membrane, the base of described filter membrane and top are individually fixed in base and the top of bag, and dual-side is individually fixed on the two sides of bag, and the width of filter membrane is less than the width of bag side; Described microbial film filtration unit comprises stainless steel stent, at least one stainless steel filter, stainless-steel vacuum table, vacuum filtration bottle, vacuum pump, wherein vacuum meter and stainless steel filter are installed on stainless steel stent by web member, and vacuum filtration bottle is housed between stainless steel stent and vacuum pump, stainless steel stent is connected by pipe connecting with vacuum filtration bottle, and described stainless steel filter comprises stainless steel base and is installed on the stainless steel funnel on stainless steel base; Described stainless steel base is provided with stainless steel filter membrane supporting pad, and stainless steel filter membrane supporting pad is suppressed by stainless steel particle and formed.
5. device according to claim 4, is characterized in that, the dual-side of described filter membrane is individually fixed in the mid-way of bag two sides.
6. device according to claim 4, is characterized in that, described stainless steel funnel is connected on stainless steel base by the mode of inlay card.
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| CN201310018933.9A CN103045713B (en) | 2013-01-18 | 2013-01-18 | Method for detecting trace amounts of microbes in a liquid sample containing multiple impurities |
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| CN201310018933.9A CN103045713B (en) | 2013-01-18 | 2013-01-18 | Method for detecting trace amounts of microbes in a liquid sample containing multiple impurities |
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| CN103045713B true CN103045713B (en) | 2015-05-27 |
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| CN105287643B (en) * | 2014-09-19 | 2018-11-27 | 平邑县人民医院 | A kind of intestinal contents component extracting |
| CN104914100B (en) * | 2015-06-04 | 2018-01-02 | 苏州大学 | The live quick visualization detection method and its kit of a kind of bacterium |
| CN106215499B (en) * | 2016-07-19 | 2018-11-23 | 浙江海洋大学 | A kind of zooplankter classified filtering device and its application |
| CN108827961A (en) * | 2018-04-25 | 2018-11-16 | 清远华新达饮品有限公司 | A kind of method of microorganism in measurement pure water |
| CN108872520A (en) * | 2018-08-14 | 2018-11-23 | 杭州绿洁水务科技股份有限公司 | A kind of water-quality test method, system and equipment |
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| US4164449A (en) * | 1977-11-03 | 1979-08-14 | J. K. And Susie L. Wadley Research Institute And Blood Bank | Surface separation technique for the detection of microbial pathogens |
| CN202044829U (en) * | 2011-03-15 | 2011-11-23 | 杭州盈天科学仪器有限公司 | Air exhaust filtering and detection device |
| CN202485935U (en) * | 2012-01-06 | 2012-10-10 | 李杜娟 | Novel sampling and homogenizing bag |
| CN202610235U (en) * | 2012-05-25 | 2012-12-19 | 杭州泰林生物技术设备有限公司 | Suction filtration device |
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2013
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US4164449A (en) * | 1977-11-03 | 1979-08-14 | J. K. And Susie L. Wadley Research Institute And Blood Bank | Surface separation technique for the detection of microbial pathogens |
| CN202044829U (en) * | 2011-03-15 | 2011-11-23 | 杭州盈天科学仪器有限公司 | Air exhaust filtering and detection device |
| CN202485935U (en) * | 2012-01-06 | 2012-10-10 | 李杜娟 | Novel sampling and homogenizing bag |
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Effective date of registration: 20190520 Address after: 310000 Room 704, Building 4, Xigang Development Center, Xihu District, Hangzhou City, Zhejiang Province Patentee after: Hangzhou Prospective Biotechnology Co., Ltd. Address before: 310013 Unit 708, Block B, Zhejiang University Science Park, Hangzhou, Zhejiang Province Patentee before: Zhang Fan |
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