CN102841058A - Detection method for number of body cells in milk - Google Patents
Detection method for number of body cells in milk Download PDFInfo
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- CN102841058A CN102841058A CN2012103132319A CN201210313231A CN102841058A CN 102841058 A CN102841058 A CN 102841058A CN 2012103132319 A CN2012103132319 A CN 2012103132319A CN 201210313231 A CN201210313231 A CN 201210313231A CN 102841058 A CN102841058 A CN 102841058A
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Abstract
The invention discloses a detection method for number of body cells in milk. The detection method is used for detecting the number of the body cells in the milk with low cost, is time-saving and labor-saving and is high in reliability. Over 95 percent of the body cells in the milk are leukocytes. Thus, the detection method for the number of the body cells in the milk comprises the following steps of: (A) dropwise adding a sample to be detected on a leukocyte filtration membrane; (B) dropwise adding a cleaning agent to the leukocyte filtration membrane; (C) dropwise adding a color developing agent to the leukocyte filtration membrane; and (D) reading color of the surface of the leukocytes and semi-quantitatively determining the number of the body cells in the milk sample by comparing with a colourimetric card, wherein the cleaning agent can be water, methanol, ethanol and acetone, the pH (Potential of Hydrogen) value is within the range of 8 to 11, and the color developing agent can be 3-acetyl indole phenol or 3-(N- toluenesulfonyl-L-alanyloxyl)-indole. The method for detecting the number of the leukocytes by using a colourimetric method is low in cost, time-saving and labor-saving and is high in reliability.
Description
Technical field
The present invention relates to the method for body cell quantity in a kind of measure milk, thereby specifically be a kind of through confirming the method for the definite body cell quantity of quantity of leucocyte in the milk.
Background technology
On agricultural, body cell all is leucocyte more than 95% in the milk, detects body cell quantity in the milk accurately, by the assay method that extensively infects as mammitis.Up to the present, the CMT method is the most general detection method.Yet CMT method labor capacity is big, has user's subjective judgement, and the false negative amount is high to accepting.Also have the automated cell counting technology in addition, in this technology, sample is diluted, and the cells of different sizes and shape obtain counting through stream of cells, costs an arm and a leg based on the instrument of fluid technique, and needs the personnel of process professional training under laboratory condition, to operate.The technological instrument that the analyser volume ratio of application nanotechnology is traditional is much little, but should technology also be in developing stage.Simultaneously, required complicated mechanical pump and the valve system of analyser makes the producer be difficult to the equipment that provides cheap.Also there is not at present suitable method to detect the increase of body cell quantity in the milk that causes by clinic mastitis.
Therefore, be necessary to develop the detection method of somatic number in a kind of milk, and with low cost, reliability is high, and labor capacity is low.
Summary of the invention
The present invention discloses the detection method of somatic number in a kind of milk, is used for low cost, and is time saving and energy saving, has somatic quantity in the detection milk of higher reliability.
Body cell all is leucocyte more than 95% in the milk, and therefore, the detection method of somatic number in the milk disclosed by the invention may further comprise the steps: the sample that (A) will detect drips on the leukoreduction filter film; (B) on the leukoreduction filter film, drip clean-out system; (C) developer is added drop-wise on the leukoreduction filter film; (D) read the color of leukocyte surface, through with the more semiquantitative definite milk sample of colorimetric card in body cell quantity.
Measure in the method for quantity of leucocyte at this, clean-out system can be water or methyl alcohol, ethanol, acetone.PH value scope is 8～11.
Measure in the method for quantity of leucocyte at this, developer can be 3-acetyl group indoxyl or 3-(N-tosyl-L-alanyl oxygen base)-indoles.
The water accepting layer that cellulose filter membrane is processed can be set below filtering membrane, be used to absorb clean-out system.
Employing utilizes the method for colourimetry detection quantity of leucocyte, and is with low cost, and time saving and energy saving, has higher reliability is arranged.
Description of drawings
Fig. 1 is the test data figure of the embodiment of the invention 5.
Embodiment
Embodiment 1 chooses milk sample to be detected, chooses and can filter leucocyte fast and can when wash-out, still keep filtering membrane Leukosorb A above that, is provided with in the bottom of filtering membrane, and the water accepting layer that cellulose filter membrane is processed is used to absorb clean-out system.Clean-out system adopts the water of 1:1 and ethanol to form, and developer is selected 3-acetyl group indoxyl for use.With milk sample to be detected, drip on filtering membrane Leukosorb A one during mensuration; On the leukoreduction filter film, drip after three minutes and form three of clean-out systems by water and the ethanol of 1:1; Developer 3-acetyl group indoxyl is added drop-wise on the leukoreduction filter film one; Read the color of leukocyte surface after five minutes, through with the more semiquantitative definite milk sample of colorimetric card in body cell quantity.
Embodiment 2 chooses milk sample to be detected, chooses and can filter leucocyte fast and can when wash-out, still keep filtering membrane Leukosorb A above that, is provided with in the bottom of filtering membrane, and the water accepting layer that cellulose filter membrane is processed is used to absorb clean-out system.Clean-out system adopts the water of 2:1 and methyl alcohol to form, and developer is selected 3-(N-tosyl-L-alanyl oxygen base)-indoles for use.With blood sample to be detected, drip on filtering membrane Leukosorb A one during mensuration; On the leukoreduction filter film, drip after four minutes and form three of clean-out systems by water and the methyl alcohol of 2:1; Developer 3-(N-tosyl-L-alanyl oxygen base)-indoles is added drop-wise on the leukoreduction filter film one; Read the color of leukocyte surface after five minutes, through with the more semiquantitative definite milk sample of colorimetric card in body cell quantity.
Embodiment 3 chooses milk sample to be detected, chooses and can filter leucocyte fast and can when wash-out, still keep filtering membrane EB above that, is provided with in the bottom of filtering membrane, and the water accepting layer that cellulose filter membrane is processed is used to absorb clean-out system.Clean-out system adopts the water of 1:1 and acetone to form, and developer is selected 3-acetyl group indoxyl for use.With urine specimen to be detected, drip on filtering membrane EB one during mensuration; On the leukoreduction filter film, drip after three minutes and form three of clean-out systems by water and the acetone of 1:1; Developer 3-acetyl group indoxyl is added drop-wise on the leukoreduction filter film one; Read the color of leukocyte surface after five minutes, through with the more semiquantitative definite milk sample of colorimetric card in body cell quantity.
Embodiment 4 chooses milk sample to be detected, chooses and can filter leucocyte fast and can when wash-out, still keep filtering membrane Leukosorb A above that.Clean-out system adopts the water of 1:5 and ethanol to form, and developer is selected 3-acetyl group indoxyl for use.With milk sample to be detected, drip on filtering membrane Leukosorb A one during mensuration; On the leukoreduction filter film, drip after two minutes and form three of clean-out systems by water and the ethanol of 1:5; Developer 3-acetyl group indoxyl is added drop-wise on the leukoreduction filter film one; Read the color of leukocyte surface after four minutes, through with the more semiquantitative definite milk sample of colorimetric card in body cell quantity.
Choosing same sample car simultaneously is sent to two different laboratories and tests with FOSSMATIC.As shown in Figure 1, the result after the result that the test of use embodiment 5 methods obtains and two laboratory datas are taken the mean, its degree of correlation performance is very outstanding (R=0.9569), except few exceptions.Cause the reason of these exception data different.Part be since milk appearance too thick (mammitis infects more serious) to such an extent as to milk appearance can not suck the suction hole in the test paper fully; Other reasons is at random.
Claims (6)
1. the detection method of somatic number in the milk is characterized in that may further comprise the steps:
The milk sample that (A) will detect drips on the leukoreduction filter film; (B) on the leukoreduction filter film, drip clean-out system; (C) developer is added drop-wise on the leukoreduction filter film; (D) read the color of leukocyte surface, through with the more semiquantitative definite milk sample of colorimetric card in body cell quantity.
2. according to the detection method of somatic number in the right 1 described milk, it is characterized in that said sample is a kind of in blood, milk liquid, urine, sweat and the saliva.
3. according to the detection method of somatic number in the right 1 described milk, it is characterized in that said clean-out system, comprise water or methyl alcohol or ethanol or acetone.
4. according to the detection method of somatic number in the right 1 described milk, it is characterized in that said clean-out system pH value scope is 8～11.
5. according to the detection method of somatic number in the right 1 described milk, it is characterized in that said developer is 3-acetyl group indoxyl or 3-(N-tosyl-L-alanyl oxygen base)-indoles.
6. according to the detection method of somatic number in the right 1 described milk, it is characterized in that below the said filtering membrane water accepting layer that cellulose filter membrane is processed being set.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103132319A CN102841058A (en) | 2012-08-30 | 2012-08-30 | Detection method for number of body cells in milk |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012103132319A CN102841058A (en) | 2012-08-30 | 2012-08-30 | Detection method for number of body cells in milk |
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| CN102841058A true CN102841058A (en) | 2012-12-26 |
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030215951A1 (en) * | 2002-05-20 | 2003-11-20 | Law Wai Tak | Method and apparatus for measuring white blood cell count |
| CN1529167A (en) * | 2003-10-10 | 2004-09-15 | 杨凌瑞特生物技术研究所 | Quick-acting diagnosis for mammitis and medicine-allergy test reagent box |
| CN1789983A (en) * | 2005-11-30 | 2006-06-21 | 浙江大学 | Method for determining somatic cell count in milk |
| CN1820198A (en) * | 2002-12-19 | 2006-08-16 | 3M创新有限公司 | Colorimetric sensors constructed of diacetylene materials |
| JP2008014688A (en) * | 2006-07-04 | 2008-01-24 | Matsushita Electric Ind Co Ltd | Somatic cell measuring and inspection kit |
| US20080057596A1 (en) * | 2006-08-30 | 2008-03-06 | Wai Tak Law | Colorimetric determination of somatic cell count in milk |
| US20100047848A1 (en) * | 2006-08-30 | 2010-02-25 | Wai Tak Law | Colorimetric determination of somatic cell count in milk |
| CN101939649A (en) * | 2008-01-07 | 2011-01-05 | 卢米耐克斯公司 | Isolation and enumeration of cells from a complex sample matrix |
| CN102279178A (en) * | 2010-06-10 | 2011-12-14 | 东北农业大学 | Method for analyzing and testing body cell content in raw milk |
-
2012
- 2012-08-30 CN CN2012103132319A patent/CN102841058A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030215951A1 (en) * | 2002-05-20 | 2003-11-20 | Law Wai Tak | Method and apparatus for measuring white blood cell count |
| CN1820198A (en) * | 2002-12-19 | 2006-08-16 | 3M创新有限公司 | Colorimetric sensors constructed of diacetylene materials |
| CN1529167A (en) * | 2003-10-10 | 2004-09-15 | 杨凌瑞特生物技术研究所 | Quick-acting diagnosis for mammitis and medicine-allergy test reagent box |
| CN1789983A (en) * | 2005-11-30 | 2006-06-21 | 浙江大学 | Method for determining somatic cell count in milk |
| JP2008014688A (en) * | 2006-07-04 | 2008-01-24 | Matsushita Electric Ind Co Ltd | Somatic cell measuring and inspection kit |
| US20080057596A1 (en) * | 2006-08-30 | 2008-03-06 | Wai Tak Law | Colorimetric determination of somatic cell count in milk |
| US20100047848A1 (en) * | 2006-08-30 | 2010-02-25 | Wai Tak Law | Colorimetric determination of somatic cell count in milk |
| CN101939649A (en) * | 2008-01-07 | 2011-01-05 | 卢米耐克斯公司 | Isolation and enumeration of cells from a complex sample matrix |
| CN102279178A (en) * | 2010-06-10 | 2011-12-14 | 东北农业大学 | Method for analyzing and testing body cell content in raw milk |
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Application publication date: 20121226 |