CN1529167A - Quick-acting diagnosis for mammitis and medicine-allergy test reagent box - Google Patents
Quick-acting diagnosis for mammitis and medicine-allergy test reagent box Download PDFInfo
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- CN1529167A CN1529167A CNA200310105811XA CN200310105811A CN1529167A CN 1529167 A CN1529167 A CN 1529167A CN A200310105811X A CNA200310105811X A CN A200310105811XA CN 200310105811 A CN200310105811 A CN 200310105811A CN 1529167 A CN1529167 A CN 1529167A
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Abstract
The invented kit provides features of fast diagnosis and detection, sensitivity, easy of operation without need of professional personnel, and on-site measurement. Bacteriostasis test for extract of Chinese traditional medicine or hydrophobicity medication shows that effect of the invented kit is better than effect of traditional paper strip method. The kit contains reagents of MTT and tetrazole triphenyl chlorinated (TTC). In detecting mastitis of milk cow following steps are carried out: adding milk sample and 0.1ml reagents of MTT and TTC at room temperature for shortest acting time 4-5 minutes; adding soluble color development liquid, and observing result. Based on whether color is appeared, the result is determined.
Description
One, technical field:
The present invention relates to a kind of mammitis quick diagnosis and drug sensitive test kit, it can be used for fast detecting and diagnosis, the test of mastitis for milk cows pathogen quick medicine-sensitive, bacterial susceptibility test, medicine bacteriostatic test and the animal products microorganism detection of cow subclinical mastitis.
Two, background technology:
The incidence of disease of cow subclinical mastitis is one of the most serious disease in cattle farm at 25%-80%.Can cause that ill milk crop reduces 20%-30%, thereby be to cause that loss is maximum, also be simultaneously to have a strong impact on one of reason that milk quality and milk cow eliminate.The antibiotic use of the main dependence of its prevention and treatment for this reason, has caused antibiotic abuse in recent years, and simultaneously, antibiotic residue has had a strong impact on the quality of milk, causes damage to the consumer, has also influenced the economic benefit of cattle farm.
The cause of disease kind that causes mammitis has reported more than 130 kinds that the pathogen feature of various places is also different.Therefore, field quick detection and drug sensitive test are significant for this disease of control.
Mainly use somatic number counting (SCC) and California mammitis test (CMT) in the milk at present in the world, China mainly uses SCC and the test of Lanzhou mammitis (LMT) and Beijing, Hebei mammitis test (BMT, HMT) etc. detect.The SCC method requires professional equipment and personnel, is difficult in plant and promotes, and various mammitises tests are then simple because of it, fast and can be used widely by execute-in-place.But said method can only carry out etiologic diagnosis, can not analyze what of microorganism wherein, more can not carry out drug sensitive test, need at first carry out separation and Culture and the purifying of bacterium at present, carry out drug sensitive test again in specialized laboratory, could instruct clinical accurate medication, separate to need a large amount of samplings owing to carry out bacterium, thereby this method is time-consuming, effort, can not execute-in-place, only regularly use in the large-scale plant of only a few at present, most of plants still rely on animal doctor's medication by rule of thumb.So far there is not to carry out the medicine box supply of on-the-spot quick medicine-sensitive test on the China market.
Three, summary of the invention:
The present invention is in order to solve the weak point in the above-mentioned background technology, a kind of mammitis quick diagnosis and drug sensitive test kit are provided, its diagnosis is quick with detection, responsive, simple to operate, need not professional and technical personnel's operation, can carry out on-site measurement, and obviously be better than conventional paper disk method for the bacteriostatic test effect of Chinese medical extract or hydrophobic drug.
For achieving the above object, the technical solution used in the present invention is:
A kind of mammitis quick diagnosis and drug sensitive test kit, its special character is: described kit contains reagent look azoles indigo plant (MTT) and chlorinated triphenyl tetrazole (TTC).
Carrying out mastitis for milk cows when detecting, add milk sample and MTT or TTC test solution 0.1mL simultaneously, room temperature or 37 ℃ the shortest 4-5 of effect minute add dissolving colour developing liquid, and observations is judged according to whether color occurring.
When carrying out the mastitis for milk cows drug sensitive test, detect positive milk sample for mammitis, add milk sample and each 0.01mL of soup, room temperature or in 37 ℃ cultivate 8-18 hour after, add MTT, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
When carrying out bacterial susceptibility test or medicine bacteriostatic test, for purified bacterium, add bacterium liquid 0.01mL, add soup 0.01mL simultaneously, after 8-18 hour, add MTT in 37 ℃ of cultivations, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
When carrying out the animal products microorganism detection, livestock products sample to be checked is pressed mixing homogenate in 1: 1 with aseptic nutrient broth, getting supernatant 0.01mL after centrifugal adds in the detection plate hole, in 37 ℃ cultivate 8-18 hour after, add MTT, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
Compared with prior art, the advantage and the effect that have of the present invention is as follows:
1, the present invention diagnosis with detect fast, sensitivity, simple to operate, need not professional and technical personnel's operation, can carry out on-site measurement, obviously be better than conventional paper disk method for the bacteriostatic test effect of Chinese medical extract or hydrophobic drug.
2, through comparative experimental research, mammitis testing result of the present invention is consistent with SCC method and LMT method.
3, mammitis drug sensitive test of the present invention and drug sensitive test testing result are consistent with microbiology conventional method (paper disk method and liquid tube method) result.
Four, embodiment:
The present invention is on the basis of multianalysis China mammitis pathogeny feature and bacterial metabolism characteristics, in conjunction with modern zymotechnic application and development, respectively with look azoles indigo plant (MTT) and chlorinated triphenyl tetrazole (TTC) substrate as the enzyme that produced in the microbial metabolism, make Bacteria Detection and drug sensitive test kit, can in 5 minutes, draw the recessive mastitis testing result, obtain the drug sensitive test result in 12-24 hour, and can be used for the cattle farm execute-in-place.
Technological process
1. mammitis detects: add milk sample and MTT or TTC test solution 0.1mL simultaneously, room temperature or 37 ℃ the shortest 4-5 of effect minute add dissolving colour developing liquid, and observations is judged according to whether color occurring.
2. mammitis drug sensitive test: detect positive milk sample for mammitis, add milk sample and each 0.01mL of soup, room temperature or in 37 ℃ cultivate 8-18 hour after, add MTT, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
3. bacterial susceptibility test: for purified bacterium, add bacterium liquid 0.01mL, add soup 0.01mL simultaneously, after 8-18 hour, add MTT in 37 ℃ of cultivations, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
4. medicine bacteriostatic test: its operating process is with 3;
5. animal products microorganism detection: livestock products sample to be checked with aseptic nutrient broth by mixing homogenate at 1: 1, getting supernatant 0.01mL after centrifugal adds in the detection plate hole, in 37 ℃ cultivate 8-18 hour after, add MTT, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
In above-mentioned each test, all can reach the animal products pollution level, and can use microplate reader and carry out detection by quantitative according to the ill degree of shade degree analyzing, medicaments insensitive degree.
Claims (5)
1, a kind of mammitis quick diagnosis and drug sensitive test kit is characterized in that: described kit contains reagent look azoles indigo plant (MTT) and chlorinated triphenyl tetrazole (TTC).
2, mammitis quick diagnosis according to claim 1 and drug sensitive test kit, it is characterized in that: when carrying out the mastitis for milk cows detection, add milk sample and MTT or TTC test solution 0.1mL simultaneously, room temperature or 37 ℃ the shortest 4-5 of effect minute, add dissolving colour developing liquid, observations is judged according to whether color occurring.
3, mammitis quick diagnosis according to claim 1 and drug sensitive test kit, it is characterized in that: when carrying out the mastitis for milk cows drug sensitive test, detect positive milk sample for mammitis, add milk sample and each 0.01mL of soup, room temperature or in 37 ℃ cultivate 8-18 hour after, add MTT, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
4, mammitis quick diagnosis according to claim 1 and drug sensitive test kit, it is characterized in that: when carrying out bacterial susceptibility test or medicine bacteriostatic test, for purified bacterium, add bacterium liquid 0.01mL, add soup 0.01mL simultaneously, in 37 ℃ cultivate 8-18 hour after, add MTT, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
5, mammitis quick diagnosis according to claim 1 and drug sensitive test kit, it is characterized in that: when carrying out the animal products microorganism detection, livestock products sample to be checked is pressed mixing homogenate in 1: 1 with aseptic nutrient broth, get supernatant 0.01mL after centrifugal and add in the detection plate hole, after 8-18 hour, add MTT in 37 ℃ of cultivations, continue to cultivate after 4 hours, add dissolving colour developing liquid, observations is judged according to whether color occurring.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200310105811XA CN1529167A (en) | 2003-10-10 | 2003-10-10 | Quick-acting diagnosis for mammitis and medicine-allergy test reagent box |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200310105811XA CN1529167A (en) | 2003-10-10 | 2003-10-10 | Quick-acting diagnosis for mammitis and medicine-allergy test reagent box |
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| CN1529167A true CN1529167A (en) | 2004-09-15 |
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| CNA200310105811XA Pending CN1529167A (en) | 2003-10-10 | 2003-10-10 | Quick-acting diagnosis for mammitis and medicine-allergy test reagent box |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100460866C (en) * | 2005-11-23 | 2009-02-11 | 山东奥克斯生物技术有限公司 | Reagent for fast diagnosing mammitis of ox, sheep, pig, its preparation process and application |
| CN101865922A (en) * | 2010-05-17 | 2010-10-20 | 北京易斯威特生物医学科技有限公司 | Milk somatic cell detection kit and method |
| CN101200755B (en) * | 2007-12-05 | 2011-06-22 | 华南农业大学 | Bacterium identification reagent kit as well as preparation method and uses thereof |
| CN102841058A (en) * | 2012-08-30 | 2012-12-26 | 山西华诚睿光生物科技有限公司 | Detection method for number of body cells in milk |
| CN103060186A (en) * | 2012-12-24 | 2013-04-24 | 山东省海水养殖研究所 | Aquatic product Edwardsiella tarda rapid drug allergy detection kit |
| CN109406345A (en) * | 2018-12-14 | 2019-03-01 | 华中农业大学 | A kind of drug sensitive test sampler and susceptibility test methods |
| CN110376366A (en) * | 2019-07-19 | 2019-10-25 | 吉林大学 | A kind of niacin is applied to the experimental method for the treatment of mastadenitis of cow by GPR109A receptor |
-
2003
- 2003-10-10 CN CNA200310105811XA patent/CN1529167A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100460866C (en) * | 2005-11-23 | 2009-02-11 | 山东奥克斯生物技术有限公司 | Reagent for fast diagnosing mammitis of ox, sheep, pig, its preparation process and application |
| CN101200755B (en) * | 2007-12-05 | 2011-06-22 | 华南农业大学 | Bacterium identification reagent kit as well as preparation method and uses thereof |
| CN101865922A (en) * | 2010-05-17 | 2010-10-20 | 北京易斯威特生物医学科技有限公司 | Milk somatic cell detection kit and method |
| CN102841058A (en) * | 2012-08-30 | 2012-12-26 | 山西华诚睿光生物科技有限公司 | Detection method for number of body cells in milk |
| CN103060186A (en) * | 2012-12-24 | 2013-04-24 | 山东省海水养殖研究所 | Aquatic product Edwardsiella tarda rapid drug allergy detection kit |
| CN109406345A (en) * | 2018-12-14 | 2019-03-01 | 华中农业大学 | A kind of drug sensitive test sampler and susceptibility test methods |
| CN109406345B (en) * | 2018-12-14 | 2024-02-09 | 华中农业大学 | Drug sensitivity test sampler and drug sensitivity test method |
| CN110376366A (en) * | 2019-07-19 | 2019-10-25 | 吉林大学 | A kind of niacin is applied to the experimental method for the treatment of mastadenitis of cow by GPR109A receptor |
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