KR100644452B1 - Fluid preservatives - Google Patents
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- KR100644452B1 KR100644452B1 KR1020040056311A KR20040056311A KR100644452B1 KR 100644452 B1 KR100644452 B1 KR 100644452B1 KR 1020040056311 A KR1020040056311 A KR 1020040056311A KR 20040056311 A KR20040056311 A KR 20040056311A KR 100644452 B1 KR100644452 B1 KR 100644452B1
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- preservative
- blood cells
- body fluid
- fluid
- edta
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- 239000003755 preservative agent Substances 0.000 title claims abstract description 27
- 239000012530 fluid Substances 0.000 title description 5
- 210000001124 body fluid Anatomy 0.000 claims abstract description 27
- 239000010839 body fluid Substances 0.000 claims abstract description 26
- 230000002335 preservative effect Effects 0.000 claims abstract description 23
- 210000000265 leukocyte Anatomy 0.000 claims abstract description 17
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 16
- 239000000203 mixture Substances 0.000 claims abstract description 4
- 239000000126 substance Substances 0.000 claims abstract description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 18
- 210000000601 blood cell Anatomy 0.000 claims description 10
- 210000002700 urine Anatomy 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 238000005070 sampling Methods 0.000 claims description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 2
- 229910021538 borax Inorganic materials 0.000 claims description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 2
- 239000004327 boric acid Substances 0.000 claims description 2
- UQGFMSUEHSUPRD-UHFFFAOYSA-N disodium;3,7-dioxido-2,4,6,8,9-pentaoxa-1,3,5,7-tetraborabicyclo[3.3.1]nonane Chemical compound [Na+].[Na+].O1B([O-])OB2OB([O-])OB1O2 UQGFMSUEHSUPRD-UHFFFAOYSA-N 0.000 claims description 2
- 239000011780 sodium chloride Substances 0.000 claims description 2
- 239000004328 sodium tetraborate Substances 0.000 claims description 2
- 235000010339 sodium tetraborate Nutrition 0.000 claims description 2
- 239000008098 formaldehyde solution Substances 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 14
- 238000005259 measurement Methods 0.000 abstract description 6
- 230000003111 delayed effect Effects 0.000 abstract description 5
- 238000004820 blood count Methods 0.000 description 16
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 206010003445 Ascites Diseases 0.000 description 3
- 239000003146 anticoagulant agent Substances 0.000 description 3
- 229940127219 anticoagulant drug Drugs 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 210000003567 ascitic fluid Anatomy 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000003748 differential diagnosis Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 210000004910 pleural fluid Anatomy 0.000 description 2
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 206010027202 Meningitis bacterial Diseases 0.000 description 1
- 206010027259 Meningitis tuberculous Diseases 0.000 description 1
- 206010027260 Meningitis viral Diseases 0.000 description 1
- 208000005228 Pericardial Effusion Diseases 0.000 description 1
- 208000022971 Tuberculous meningitis Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 210000004381 amniotic fluid Anatomy 0.000 description 1
- 201000009904 bacterial meningitis Diseases 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 208000001223 meningeal tuberculosis Diseases 0.000 description 1
- 238000012009 microbiological test Methods 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 210000004912 pericardial fluid Anatomy 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 201000010044 viral meningitis Diseases 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/10—Preservation of living parts
- A01N1/12—Chemical aspects of preservation
- A01N1/122—Preservation or perfusion media
- A01N1/126—Physiologically active agents, e.g. antioxidants or nutrients
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
본 발명은 임상 검체 중, 체액내에서 백혈구 및 적혈구수의 측정을 위한 검체의 보존제에 관한 것이다.The present invention relates to a preservative of a sample for the measurement of leukocytes and erythrocyte counts in body fluids in clinical samples.
본 발명은 백혈구 및 적혈구의 변형을 방지하기 위한 화학적 조성물로서 검체 채취 후, 본 화학적 조성물과 동량을 혼합하여 보관하면 백혈구 및 적혈구수의 변화가 적어 검사가 지연될 경우에 정확한 검사가 가능하다.The present invention is a chemical composition for preventing the deformation of white blood cells and red blood cells, and after the sample is collected, if the chemical composition and the same amount is stored and stored, the changes in the number of white blood cells and red blood cells are small, so that accurate testing can be performed when the test is delayed.
체액, 적혈구수, 백혈구수, 측정, 보존제Body fluid, erythrocyte count, leukocyte count, measurement, preservative
Description
임상검체 중 뇌척수액을 포함한 흉수, 복수, 심낭액, 양수, 관절액, 요 등에서 적혈구 및 백혈구(호중구, 호산구, 림프구)수의 측정은 질병의 진단, 치료후 관찰 및 예후 판정에서 임상적으로 중요하다. 특히 뇌척수액내에서 백혈구수의 측정은 결핵성 수막염, 세균성 수막염 및 바이러스 수막염의 감별진단에 이용되어 긴급한 감별진단과 치료에서 중요한 검사항목으로 백혈구수의 정확한 측정은 매우 중요하다.The measurement of erythrocytes and leukocytes (neutrophils, eosinophils, lymphocytes) in cerebrospinal fluid, ascites, pericardial fluid, amniotic fluid, joint fluid, and urine, including cerebrospinal fluid, is clinically important in the diagnosis of disease, post-treatment observation, and prognosis. In particular, the measurement of white blood cell count in cerebrospinal fluid is used for differential diagnosis of tuberculous meningitis, bacterial meningitis and viral meningitis. Therefore, it is very important test item for urgent differential diagnosis and treatment.
현재 체액내의 백혈구 및 적혈구수의 측정은 체액내의 혈구의 응집을 억제하기 위하여 검체를 K3EDTA, Heparin 등의 항응고제가 포함된 시험관에 채취한 후, 시산법을 이용하여 세포수를 계수한다. 시산법은 수동혈구측정기를 이용하여 세포수를 계수하는데, 수동혈구측정기는 유리 또는 플라스틱 재질로 제작되었으며 검체가 모세관 현상에 의해 확산되어 일정한 양이 들어갈 수 있도록 일정한 부피의 공간으로 이루어져 이를 직접 광학현미경하에서 관찰하여 혈구수를 계수하여 혈구수를 산정한다. 시산법에서는 검체의 시간 경과에 따라 세포의 변형이 일어나 모양 판별이 어려운 경우가 많아 측정자의 숙련도에 따라 차이가 난다. 이와같이 검체를 채취한 후, 시간이 경과할수록 각 혈구의 변형이 일어나 혈구의 감별이 어렵고, 측정된 백혈구 및 적혈구 수는 감소하며, 특히 체액의 경우에는 다른 미생물학적 검사를 시행하기 위해 실온에 보관하는데 이러한 경우 감소율은 더 증가한다.To measure the white blood cell and red blood cell count in the body fluid, the sample is collected in a test tube containing anticoagulant such as K 3 EDTA, Heparin, and counted the cell number in order to suppress aggregation of blood cells in the body fluid. The counting method uses a manual hemocytometer to count cell numbers. The manual hemocytometer is made of glass or plastic material and consists of a volume of space so that the sample can be diffused by capillary action and enter a certain amount. Observed below to count blood counts to calculate blood counts. In the trial method, it is difficult to distinguish the shape due to the deformation of the cell with the passage of time of the sample, and the difference depends on the skill of the measurer. As the sample is collected, the blood cells are deformed over time, making it difficult to differentiate the cells, and the measured number of white blood cells and red blood cells decreases, especially in the case of body fluids, stored at room temperature for other microbiological tests. In this case, the reduction rate is further increased.
일반적으로 병원에서 체액내의 혈구수의 측정은 30분이내에 측정하도록 되어 있는데 실제로 검체 채취후 시간의 경과에 따라 혈구수는 감소하며, 검사자가 즉시 측정이 불가능하거나 자체기관에 검사가 불가능하여 수탁기관으로 검사를 의뢰하는 등, 검사가 지연될 경우 및 검사 결과가 이상하여 재검을 할 경우에는 정확한 혈구수의 측정이 어려워 질병의 진단이 잘못되는 경우가 있으므로 체액내에서의 각 혈구수의 정확한 측정을 위해서는 검체채취 후, 즉시 측정해야 한다.In general, blood cell counts in body fluids are measured within 30 minutes in hospitals. Actually, blood cell counts decrease with the passage of time after the sample is collected. If the test is delayed, such as requesting a test, or if the test result is abnormal, and the test is performed again, accurate blood counts may be difficult to diagnose, and the diagnosis of the disease may be wrong. After sampling, it should be measured immediately.
본 발명의 목적은 임상진단시 이용되는 체액내의 백혈구 및 적혈구수 측정을 위해 검체 채취 후, 검사가 지연되어도 정확한 결과를 얻을수 있으며, 재검시에도 상대적으로 정확한 결과를 얻을 수 있도록 각 혈구의 모양이 변형되지 않고 측정시 각 혈구의 모양이 잘 관찰되도록 하는 보존제를 이용하여 정확하게 측정할 수 있는 방법을 제공하는 것이다.
The purpose of the present invention is to obtain accurate results even if the test is delayed after sampling for measuring white blood cells and red blood cell count in the body fluids used in clinical diagnosis, and the shape of each blood cell is modified to obtain a relatively accurate result even when retesting. It is to provide a method that can be accurately measured using a preservative to ensure that the shape of each blood cell is observed at the time of measurement.
상기 목적을 달성하기 위하여, 본 발명은 체액 채취시에 세포의 모양이 변형되지 않는 보존제를 검체와 동량으로 혼합하여 보관하고 혼합된 검체를 이용하여 각 혈구수를 측정한 후, 세포수 측정값에 희석배수인 1/2을 곱하여 혈구수를 알 수 있다. 기존의 항응고제만을 이용한 혈구수 측정 방법의 경우, 검사가 지연될 경우 및 추후 재확인시에는 각 혈구의 모양이 변형되므로 측정된 혈구수의 값이 감소하여 측정값이 부정확하였으나 검체 보존제를 혼합함으로써 정확한 측정값을 얻을 수 있도록 하는 특징을 가지고 있다.
In order to achieve the above object, the present invention is to store and store the same amount of preservatives that do not change the shape of the cell at the time of body fluid collection and the same amount of the sample, and measure the number of blood cells using the mixed sample, The blood count can be obtained by multiplying the dilution factor 1/2. In the case of blood count measurement method using only anticoagulant, the shape of each blood cell is deformed when the test is delayed and later rechecked. It has a feature that allows you to get a value.
본 발명은 백혈구 및 적혈구의 변형을 방지하여 시산법으로 혈구 계수시 혈구의 판별이 용이하며 정확한 측정이 가능하도록 하기 위한 보존제로 pH, 오스몰농도(osmolarity)를 최적화하고 항응고제 및 고정제(fixative)를 첨가하여 체액내의 각 혈구의 변형을 방지할 수 있는 완충액(buffer)으로 조성된 화학적 조성물로 각 성분 및 농도는 아래와 같다.The present invention optimizes pH, osmolarity, and anticoagulants and fixatives as a preservative to prevent the deformation of white blood cells and red blood cells, so that blood cells can be easily identified and accurately measured by counting methods. The chemical composition composed of a buffer (buffer) to prevent the deformation of each blood cell in the body fluid by adding the components and concentrations are as follows.
1. Sodium chloride 6.38 g/LSodium chloride 6.38 g / L
2. Boric acid 1.0 g/LBoric acid 1.0 g / L
3. Sodium tetraborate 0.2 g/L3.Sodium tetraborate 0.2 g / L
4. K3 EDTA 0.2 g/LK 3 EDTA 0.2 g / L
5. Formaldhyde solution(37%) 10 ml/L 5.Formaldhyde solution (37%) 10 ml / L
본 체액 보존제를 검체 채취시 동량의 검체와 보존제를 혼합하여 혼합하여 보관하고 혼합된 검체를 이용하여 각 혈구수를 측정한 후, 세포수 측정값에 희석배 수인 1/2을 곱하여 정확한 혈구수를 측정할 수 있다. 이와같이 본 체액 보존제를 이용함으로써 알 수 있다.When the sample is collected, the same amount of sample and preservative are mixed and stored, and each blood cell number is measured using the mixed sample, and the cell count is multiplied by 1/2 the dilution factor to obtain an accurate blood count. It can be measured. Thus, it can know by using this body fluid preservative.
이하, 본 발명의 실시예는 다음과 같다.Hereinafter, embodiments of the present invention are as follows.
백혈구 및 적혈구가 발견되는 환자의 뇌척수액, 흉수, 복수 및 요 각각 10예를 대상으로 기존의 K3 EDTA가 함유되어 있는 시험관 및 본 체액보존제가 포함되어 있는 시험관에 각각 검체를 채취하였고, 각각을 검체 채취 후, 1시간이내, 2시간, 4시간 및 12시간째에 Naegeotte chamber를 이용한 수기법으로 혈구수를 측정하여 비교하였다. 모든(뇌척수액, 흉수, 복수 및 요) 검체에서 검체 채취 후 즉시 검사한 결과, 기존의 K3 EDTA 시험관 및 본 체액보존제가 포함된 검체 간에 백혈구 및 적혈구의 유의한 차이는 보이지 않았으나(p<0.05), 기존의 K3 EDTA 시험관내의 검체에서 비교적 낮게(표1) 측정 되었다. Ten cases of cerebrospinal fluid, pleural fluid, ascites, and urine in patients with leukocytes and erythrocytes were found in 10 test tubes containing K 3 EDTA and test tubes containing body fluid preservatives, respectively. Within 1 hour, 2 hours, 4 hours, and 12 hours after collection, blood cell counts were measured by a manual method using a Naegeotte chamber. Immediately after sampling from all (cerebrospinal fluid, pleural fluid, ascites and urine) specimens, no significant differences in leukocytes and erythrocytes were observed between conventional K 3 EDTA test tubes and samples containing body fluid preservatives ( p <0.05). The results were relatively low in conventional K 3 EDTA in vitro specimens (Table 1).
시간의 경과에 따라 K3 EDTA 및 본 체액보존제를 이용하여 백혈구 및 적혈구수를 측정한 결과, 시간이 경과하면 각 검체에서 백혈구수 및 적혈구수가 감소하였으며, 본 체액보존제를 이용하는 경우가 기존의 K3 EDTA를 이용하여 측정하는 경우보다 변화가 적었다.(표2, 표3)Over a period of time K 3 EDTA and the results of the measurement of the white blood cells and the number of red blood cells using the body fluid preservative, over time, decreased the number and red blood cell number of white blood cells in each sample, the existing K 3 when using the present fluid preservative The change was smaller than that measured using EDTA (Tables 2 and 3).
* 초기 측정값을 100%로 하였을 때의 %값.(평균±표준편차)*% Value when initial measured value is 100% (mean ± standard deviation)
* 초기 측정값을 100%로 하였을 때의 %값.(평균±표준편차)*% Value when initial measured value is 100% (mean ± standard deviation)
이상과 같이 본 발명은 체액내의 백혈구 및 적혈구의 변형을 방지하여 검사가 지연될때 혈구수의 감소를 방지하고 수기법으로 혈구수 측정시 관찰이 용이하도록 해주는 체액용 보존제로서 이를 이용하면 즉시 검사하기 어려운 경우나 재검시에 용이하게 혈구수를 정확히 측정할 수 있다.As described above, the present invention is a preservative for bodily fluids that prevents the decrease of blood cell count when the test is delayed by preventing deformation of white blood cells and red blood cells in the body fluid, and makes it easy to observe when measuring blood cell count by manual technique. Blood counts can be accurately measured at the time of re-examination or examination.
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| GB0704488D0 (en) * | 2007-03-08 | 2007-04-18 | Univ Nottingham | Platelet viability kit |
| US9321664B2 (en) | 2011-12-20 | 2016-04-26 | Ecolab Usa Inc. | Stable percarboxylic acid compositions and uses thereof |
| US10165774B2 (en) | 2013-03-05 | 2019-01-01 | Ecolab Usa Inc. | Defoamer useful in a peracid composition with anionic surfactants |
| KR102194417B1 (en) * | 2019-01-30 | 2020-12-24 | (주)메디젠휴먼케어 | A composition for preserving body fluids, and a preparation method thereof |
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| US4258032A (en) | 1979-01-15 | 1981-03-24 | Becton, Dickinson And Company | Preservation of urine specimens |
| JPH02129127A (en) * | 1988-11-04 | 1990-05-17 | Sumitomo Seika Chem Co Ltd | Improved red blood cell preservation solution |
| JPH08201382A (en) * | 1995-01-24 | 1996-08-09 | Nippon Kayaku Co Ltd | Analyzing method for body fluid component and body fluid collection sheet |
| JPH10265769A (en) | 1997-03-26 | 1998-10-06 | Asahi Denka Kogyo Kk | Soft gel-like cooling storage material |
| JP2002080314A (en) | 2000-06-19 | 2002-03-19 | Santen Pharmaceut Co Ltd | Preservative |
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- 2004-07-20 KR KR1020040056311A patent/KR100644452B1/en active IP Right Grant
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4258032A (en) | 1979-01-15 | 1981-03-24 | Becton, Dickinson And Company | Preservation of urine specimens |
| JPH02129127A (en) * | 1988-11-04 | 1990-05-17 | Sumitomo Seika Chem Co Ltd | Improved red blood cell preservation solution |
| JPH08201382A (en) * | 1995-01-24 | 1996-08-09 | Nippon Kayaku Co Ltd | Analyzing method for body fluid component and body fluid collection sheet |
| JPH10265769A (en) | 1997-03-26 | 1998-10-06 | Asahi Denka Kogyo Kk | Soft gel-like cooling storage material |
| JP2002080314A (en) | 2000-06-19 | 2002-03-19 | Santen Pharmaceut Co Ltd | Preservative |
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| KR20060007497A (en) | 2006-01-26 |
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