CN107589251A - Lupus anticoagulant analyte detection kit and lupus anticoagulant presence or absence determination methods - Google Patents
Lupus anticoagulant analyte detection kit and lupus anticoagulant presence or absence determination methods Download PDFInfo
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Abstract
The method of lupus anticoagulant presence or absence in testing sample, including the 1st time of setting test reagent are judged the invention discloses lupus anticoagulant analyte detection kit and by the kit, it includes clotting factor activator, soybean lecithin, CaCl2And buffer solution；2nd time of setting test reagent, it includes clotting factor activator, hydrogenated phospholipid, CaCl2And buffer solution；The present invention adds hydrogenated phospholipid in the 2nd time of setting test reagent, and with hydrogenated phospholipid combination LA, specificity is good, and stability is high；The kit of the present invention is when detecting LA, other supporting known detection reagents are not needed, without the time of setting test reagent of the 1st time of setting test reagent/the 2nd and healthy normal plasma are mixed to carry out the judgement of positive sample, and simplify testing process without sample process is carried out before test sample.
Description
Technical field
The present invention relates to lupus anticoagulant detection technique field, more particularly to a kind of lupus anticoagulant analyte detection kit,
The invention further relates to the method for judging lupus anticoagulant presence or absence in test plasma.
Background technology
Lupus anticoagulant (lupus anticoagulation, LA) is a kind of for itself resisting with negative charge phosphatide
Body, it is one kind of anti-phospholipid antibody, is common in the connective tissue Disease such as systemic loupus erythematosus.Because it is first in erythema
It is studied with lupus patient, therefore it is named as Lupus anticoagulant.Have now found that it may be present in a variety of diseases.Lupus anticoagulant
To exist be considered as the habitual abortion of unknown cause, stillborn foetus, development of fetus sluggishness, arteriovenous embolism, various for material lasting
The danger signal of easy bolt disease and some autoimmune diseases.
LA can block the clotting factor and fibrin ferment original work of activation by identifying that fat bind thrombin influences Coagulation test originally
With, so as to suppress fibrinous formation, cause cruor time extending, therefore referred to as antiprothrombin antibody be likely more it is suitable
Preferably, carry out medical diagnosis on disease of the LA experiment detection to clinical departments to have a very big significance.
LA detection includes Screening tests, bulk testing and validation test.Because LA influence factor is complicated, in Screening tests rank
Section is, it is necessary to by prothrombin time（PT）, fibrinogen detection, dilution or improvement after Russell venom times（dRVVT
Or MRVVT) and Staclot-LA experiments or kaolin clotting time etc. be used in combination, to avoid each disturbing factor, but more
The combination of kind method causes operating process cumbersome.
Bulk testing is to be mixed by using healthy normal plasma and test plasma using 1: 1 ratio, to observe extension
Activated partial thromboplastin time（APTT）Whether it is repaired.If test plasma lacks one or more clotting factor, after mixing
The APTT of extension will be corrected；If the anticoagulant substanceses such as LA be present in test plasma on the contrary, can not be repaired.
On detection ordering, International Society on Thrombosis and Homeostasis（ISTH）Advocate at once to enter during the LA Screening tests positives
Row bulk testing, decide whether further to do LA validation test according to the result of bulk testing.
LA validation test is solidifying containing congealed fat with time of setting test reagent of the low concentration containing congealed fat and high concentration respectively
Gu timing reagent determines setting time, the ratio of setting time is obtained by each reagent to confirm LA in sample to be tested
In the presence of.
Authorization Notice No. CN102650643B discloses one kind, and " lupus anticoagulant analyte detection is deposited with kit and lupus anticoagulant
Whether determination methods ", including（i）1st time of setting test reagent, its contain selected from phosphatidyl-ethanolamine, phosphatidyl choline and
At least one kind of phosphatide in phosphatidylserine；And（ii）2nd time of setting test reagent, it contains selected from phosphatidyl-ethanolamine, phosphorus
At least one kind of phosphatide in phosphatidylcholine and phosphatidylserine；Wherein the 1st time of setting test reagent and the 2nd setting time are surveyed
Determine at least one party's alkali metal containing salt among reagent, and the 2nd time of setting test reagent contains and surveyed compared to the 1st setting time
Determine the alkali metal salt of reagent lower concentration, or, without alkali metal salt.
Authorization Notice No. CN102384979B discloses the " judgement of lupus anticoagulant analyte detection kit and its presence or absence
Method ", including the 1st time of setting test reagent, its manganese containing salt, phosphatide and selected from ellagic acid, kaolin, diatomite and oxidation
The activator for being used to cause in vitro blood clotting of silicon；2nd time of setting test reagent, its manganese containing salt, phosphatide and selected from tan
The activator for being used to cause in vitro blood clotting of acid, kaolin, diatomite and silica is spent, wherein, the 2nd setting time is surveyed
Determine low or the 2nd time of setting test reagent in the time of setting test reagent of manganese salt concentration ratio the 1st in reagent not manganese containing salt；
With the 3rd time of setting test reagent, it contains calcium salt.
Above two LA detections kit more clearly distinguishes LA positives testing sample and LA is cloudy than conventional kit
Property testing sample.But the kit needs to detect sample to be tested simultaneously and healthy normal sample obtained lupus ratio originally, increase
The workload of test.
The content of the invention
Need to solve LA detections kit in the prior art while detect sample to be tested and healthy normal sample is original
Obtain lupus ratio, cause the problem of workload increase, it is an object of the invention to provide a kind of lupus anticoagulant analyte detection reagent
Box.Therefore, the present invention also provides a kind of method that lupus anticoagulant presence or absence in test plasma is judged by the kit.
To achieve these goals, the present invention adopts the following technical scheme that：
The first aspect of the present invention, there is provided a kind of lupus anticoagulant analyte detection kit, including
1st time of setting test reagent, it includes clotting factor activator, soybean lecithin, CaCl2And buffer solution；
2nd time of setting test reagent, it includes clotting factor activator, hydrogenated phospholipid, CaCl2And buffer solution.
Preferably, 2-10mg/L clotting factor activator, 5- is included in the 1st time of setting test reagent
20mg/L soybean lecithin, 10-50mmol/L CaCl2With 20-80mmol/L pH=7-8 buffer solution；During the 2nd solidification
Between determine in reagent comprising 2-10mg/L clotting factor activator, 10-40mg/L hydrogenated phospholipid, 10-50mmol/L
CaCl2With 20-80mmol/L pH=7-8 buffer solution.
Preferably, the clotting factor activation in the 1st time of setting test reagent and the 2nd time of setting test reagent
Agent is RVV-X, and buffer solution is trishydroxymethylaminomethane-hydrochloric acid solution（Tris-HCl）.
It is further preferred that also include albumen in the 1st time of setting test reagent and the 2nd time of setting test reagent
Protective agent, preservative, ionic strength adjustor and clotting factor protective agent.
Preferably, 22- is also included respectively in the 1st time of setting test reagent and the 2nd time of setting test reagent
70g/L protein protective agents, 0.03-0.05% preservatives, 80-300mmol/L ionic strength adjustors and 1-6g/L clotting factor are protected
Agent is protected, above-mentioned percentage is percentage by volume.
Preferably, the protein protective agent is the compound of component A and B component, and the component A is BSA（Ox blood is pure
Albumen）, casein or skimmed milk power, the B component be trehalose, mannitol or sucrose；Described preservative is
Proclin300, described ionic strength adjustor are NaCl, KCl or MgCl2, described clotting factor protective agent is sweet ammonia
Acid, histidine or arginine.
Preferably, the component A and the compounding of 12-40g/L B component composition that the protein protective agent is 10-30g/L
Thing.
The second aspect of the present invention, there is provided the determination methods of lupus anticoagulant presence or absence are carried out using mentioned reagent box,
Comprise the following steps：
S1, >=30 people's health normal plasmas are mixed with the 1st time of setting test reagent, the 2nd time of setting test reagent respectively,
Obtain the setting time scope T1 of normal plasma the 1st, the setting time of normal plasma the 1st and the 2nd setting time ratio range T2；
S2, the test plasma gathered from subject is divided into two parts；
S3, one of will be divided into two parts in S2 and the 1st time of setting test reagent mixing determines the 1st setting time；
S4, according to the 1st setting time and T1 relation, judge whether testing sample contains lupus anticoagulant.
When the 1st setting time that test plasma measures falls into T1 scopes, it is healthy normal plasma to show test plasma；When
When the 1st setting time that test plasma measures exceedes T1 maximum, it is probable positive blood plasma to show test plasma.
Preferably, also include after the S4 steps：
S5, other a and the 2nd time of setting test reagent being divided into two parts in S2 is mixed to determine the 2nd setting time；
S6, judge whether above-mentioned test plasma contains lupus anticoagulant with the ratio of the 1st setting time and the 2nd setting time.
Test plasma is subjected to S5, S6 step, when the ratio in S6 is when between 0.9-1.2, shows that test plasma is
Feminine gender, without lupus anticoagulant；When ratio is more than 1.2, show that test plasma for the positive, contains lupus anticoagulant.
Compared with prior art, the beneficial effect that the present invention realizes：The present invention adds in the 2nd time of setting test reagent
Hydrogenated phospholipid, with hydrogenated phospholipid combination LA, specificity is good, and stability is high；The kit of the present invention is when detecting LA, it is not necessary to matches somebody with somebody
Other known detection reagents are covered, without the time of setting test reagent of the 1st time of setting test reagent/the 2nd and health is normal
Blood plasma is mixed to carry out the judgement of positive sample, and simplifies detection stream without sample process is carried out before test sample
Journey, reduce the workload of test.
Embodiment
Embodiment 1
Lupus anticoagulant analyte detection includes the 1st time of setting test reagent and the 2nd time of setting test reagent with kit.
The preparation of 1st time of setting test reagent：
S1, weigh 0.605g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 20 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.1；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, it is 1g/L to obtain RVV-X Stock concentrations；
S3, weigh 5mg soybean lecithins and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
Soybean lecithin Stock concentrations are 1g/L；
S4, weigh 0.222g CaCl2, 0.467g NaCl, 0.2g glycine, 1.0g BSA, 1.5g trehaloses, add 100ml
Volumetric flask；
S5, draws 0.2ml RVV-X storing solutions, and 0.5ml soybean lecithin storing solutions are separately added into S4 100ml volumetric flasks；
S6, draw 30ul Proclin300 and add in S5 volumetric flask, it is equal to graduation mark, mixing to add Tris-HCl solution
It is even；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
The preparation of 2nd time of setting test reagent：
S1, weigh 0.605g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 20 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.1；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, Stock concentrations 1g/L；
S3, weigh 5mg hydrogenated phospholipids and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
It is 1g/L to hydrogenate congealed fat Stock concentrations；
S4, weigh 0.222g CaCl2, 0.467g NaCl, 0.2g glycine, 1.0g BSA, 1.5g trehaloses, add 100ml
Volumetric flask；
S5, draws 0.2ml RVV-X storing solutions, and 1ml hydrogenation congealed fat storing solutions are separately added into S4 100mL volumetric flasks；S6,
Draw 30ul Proclin300 to add in S5 volumetric flask, add Tris-HCl solution to graduation mark, be well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
Embodiment 2
Lupus anticoagulant analyte detection includes the 1st time of setting test reagent and the 2nd time of setting test reagent with kit.
The preparation of 1st time of setting test reagent：
S1, weigh 1.514g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 50 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.5；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, it is 1g/L to obtain RVV-X Stock concentrations；
S3, weigh 5mg soybean lecithins and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
Soybean lecithin Stock concentrations are 1g/L；
S4, weigh 0.333g CaCl2, 0.877g NaCl, 0.4g glycine, 2.0g BSA, 2.5g trehaloses, add 100ml
Volumetric flask；
S5, draws 0.5ml RVV-X storing solutions, and 1.5ml soybean lecithin storing solutions are separately added into S4 100ml volumetric flasks；
S6, the volumetric flask that 50ul Proclin300 add S5 is drawn, add Tris-HCl solution to graduation mark, be well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
The preparation of 2nd time of setting test reagent：
S1, weigh 1.514g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 50 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.5；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, Stock concentrations 1g/L；
S3, weigh 5mg hydrogenated phospholipids and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
It is 1g/L to hydrogenate congealed fat Stock concentrations；
S4, weigh 0.333g CaCl2, 0.877g NaCl, 0.4g glycine, 2.0g BSA, 2.5g trehaloses, add 100ml
Volumetric flask；
S5, draws 0.5ml RVV-X storing solutions, and 2.5ml hydrogenation congealed fat storing solutions add volumetric flask；
S6, the volumetric flask that 50ul Proclin300 add S5 is drawn, add Tris-HCl solution to graduation mark, be well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
Embodiment 3
Lupus anticoagulant analyte detection includes the 1st time of setting test reagent and the 2nd time of setting test reagent with kit.
The preparation of 1st time of setting test reagent：
S1, weigh 2.118g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 70 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.9；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, it is 1g/L to obtain RVV-X Stock concentrations；
S3, weigh 5mg soybean lecithins and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
Soybean lecithin Stock concentrations are 1g/L；
S4, weigh 0.5549g CaCl2, 1.695g NaCl, 0.6g glycine, 3.0g BSA, 3.8g trehaloses, add 100ml
Volumetric flask；
S5, draws 1ml RVV-X storing solutions, and 2ml soybean lecithin storing solutions are separately added into S4 100ml volumetric flasks；
S6, the volumetric flask that 35ul Proclin300 add S5 is drawn, add Tris-HCl solution to graduation mark, be well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
The preparation of 2nd time of setting test reagent：
S1, weigh 2.421g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 80 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.9；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, Stock concentrations 1g/L；
S3, weigh 5mg hydrogenated phospholipids and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
It is 1g/L to hydrogenate congealed fat Stock concentrations；
S4, weigh 0.5549g CaCl2, 1.695g NaCl, 0.6g glycine, 3.0g BSA, 3.8g trehaloses, add 100ml
Volumetric flask；
S5, draws 1ml RVV-X storing solutions, and 4ml hydrogenation congealed fat storing solutions add volumetric flask；
S6, the volumetric flask that 35ul Proclin300 add S5 is drawn, add Tris-HCl solution to graduation mark, be well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
Embodiment 4
Lupus anticoagulant analyte detection includes the 1st time of setting test reagent and the 2nd time of setting test reagent with kit.
The preparation of 1st time of setting test reagent：
S1, weigh 2.421g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 80 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.6；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, it is 1g/L to obtain RVV-X Stock concentrations；
S3, weigh 5mg soybean lecithins and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
Soybean lecithin Stock concentrations are 1g/L；
S4, weigh 0.111g CaCl2, 1.753g NaCl, 0.1g glycine, 1.5g BSA, 4g trehaloses, add
100ml volumetric flasks；
S5, draws 0.8ml RVV-X storing solutions, and 1.8ml soybean lecithin storing solutions are separately added into S4 100ml volumetric flasks；
S6, the volumetric flask that 40ul Proclin300 add S5 is drawn, add Tris-HCl solution to graduation mark, be well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
The preparation of 2nd time of setting test reagent：
S1, weigh 2.421g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 80 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.6；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, Stock concentrations 1g/L；
S3, weigh 5mg hydrogenated phospholipids and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
It is 1g/L to hydrogenate congealed fat Stock concentrations；
S4, weigh 0.111g CaCl2, 1.753g NaCl, 0.1g glycine, 1.5g BSA, 4g trehaloses, add
100ml volumetric flasks；
S5, draws 0.8ml RVV-X storing solutions, and 3.5ml hydrogenation congealed fat storing solutions add volumetric flask；
S6, the volumetric flask that 40ul Proclin300 add S5 is drawn, add Tris-HCl solution to graduation mark, be well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
Embodiment 5
Lupus anticoagulant analyte detection includes the 1st time of setting test reagent and the 2nd time of setting test reagent with kit.
The preparation of 1st time of setting test reagent：
S1, weigh 0.605g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 20 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.1；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, it is 1g/L to obtain RVV-X Stock concentrations；
S3, weigh 5mg soybean lecithins and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
Soybean lecithin Stock concentrations are 1g/L；
S4, weigh 0.222g CaCl2, add 100ml volumetric flasks；
S5, draws 0.2ml RVV-X storing solutions, and 0.5ml soybean lecithin storing solutions are separately added into S4 100ml volumetric flasks, added
Enter Tris-HCl solution to graduation mark, be well mixed；
S6, every bottle of 4ml packing, 2~8 DEG C of preservations.
The preparation of 2nd time of setting test reagent：
S1, weigh 0.605g tri-（Methylol）Aminomethane（Tris）250ml volumetric flasks are put into, add distilled water, are added dense
HCl adjusts pH value, adds water to graduation mark, is well mixed, and prepares 20 mmol/L Tris-HCl solution, measuring pH with pH meter is
7.1；
S2, draw 1ml distilled water and redissolve RVV-X freeze-dried powders, Stock concentrations 1g/L；
S3, weigh 5mg hydrogenated phospholipids and add 5ml Tris-HCl solution, be ground to emulsus, visually observe without obvious particle, obtain
It is 1g/L to hydrogenate congealed fat Stock concentrations；
S4, weigh 0.222g CaCl2, add 100ml volumetric flasks；
S5,0.2ml RVV-X storing solutions are drawn, 1ml hydrogenation congealed fat storing solutions are separately added into S4 100mL volumetric flasks,
Tris-HCl solution is added to graduation mark, is well mixed；
S7, every bottle of 4ml packing, 2~8 DEG C of preservations.
Embodiment 6
By taking kit prepared by embodiment 2 as an example, testing sample is detected using Japanese SYSMEX CA1500 Automatic coagulometers,
Process is as follows：
The 1st, 1st time of setting test reagent and the 2nd time of setting test reagent are put into the reagent position set in instrument；
2nd, test plasma is positioned over sample rack；
3rd, 100ul test plasmas are taken to add the time of setting test reagents of 100ul the 1st after 37 DEG C of pre-temperature 4min, pre-temperature, measure the
1 setting time, when the 1st setting time falls into normal plasma setting time term of reference, show that test plasma resists without lupus
Condensate；When the 1st setting time exceeds normal plasma setting time term of reference, need to be carried out with the 2nd time of setting test reagent
Confirm；
4th, 100ul identicals test plasma is taken to add the time of setting test reagents of 100ul the 2nd after 37 DEG C of pre-temperature 4min, pre-temperature,
Determine the 2nd setting time；
5th, the ratio of the 1st setting time and the 2nd setting time is calculated, is compared with normal plasma setting time ratio term of reference
Compared with, judge whether lupus anticoagulant the positive.
The determination of normal plasma setting time term of reference：The healthy individuals blood plasma at 40 ages 18-55 year is collected, according to
Above-mentioned detection method, the plasma coagulation time of each sample is determined with the 1st time of setting test reagent of the present invention, what is measured is normal
Plasma coagulation time term of reference is：34-45s.
The determination of normal plasma setting time ratio term of reference：Determined with the 2nd time of setting test reagent of the present invention
2nd setting time of above-mentioned 40 healthy normal plasmas, with the 1st setting time and the 2nd setting time phase of healthy normal plasma
Than it is 0.9-1.2 to obtain normal plasma setting time ratio term of reference.
Kit of the present invention and using the kit judge test plasma whether lupus anticoagulant the positive process, not only fit
For SYSMEX CA1500 coagulo meters, but also suitable for other brands, the coagulo meter of model, such as SYSMEX other models
Coagulo meter, U.S.'s Beckman Kurt ACL Advance Automatic coagulometers, France think to reach the high full-automatic blood of STA Compact
Solidifying instrument etc..
Embodiment 7
With kit prepared by embodiment 2 and the fertile sweet smell of import（IL）LA detection reagents are carried out to clinical 200 plasma samples respectively
LA screening test, the incidence that statistics plasma coagulation time extends, to investigate the clinical specificity of reagent, as a result such as the institute of table 1
Show.
Table 1
As can be seen from Table 1, the clinical specificity of kit of the present invention reaches import reagent standard.
Embodiment 8
Kit prepared by embodiment 2 is placed in 37 DEG C of preservations, takes out during use, the same normal blood of LA is determined in different time
Slurry, the setting time of LA abnormal plasmas, as a result as shown in table 2.
Table 2
Such as table 2, kit METHOD FOR CONTINUOUS DETERMINATION of the present invention 15 days, stabilization is as a result always maintained at, it is preferable to illustrate that kit of the present invention has
Stability.
Above-mentioned embodiment is exemplary, is to preferably make skilled artisans appreciate that originally
Patent, it is impossible to be not understood as the limitation for including scope to this patent；As long as times made of spirit according to disclosed in this patent
How with change or modification, the scope that this patent includes is each fallen within.
Claims (8)
1. lupus anticoagulant analyte detection kit, it is characterised in that including
1st time of setting test reagent, it includes clotting factor activator, soybean lecithin, CaCl2And buffer solution；
2nd time of setting test reagent, it includes clotting factor activator, hydrogenated phospholipid, CaCl2And buffer solution.
2. lupus anticoagulant analyte detection kit as claimed in claim 1, it is characterised in that the 1st time of setting test
2-10mg/L clotting factor activator, 5-20mg/L soybean lecithin, 10-50mmol/L CaCl are included in reagent2And 20-
80mmol/L pH=7-8 buffer solution；The clotting factor comprising 2-10mg/L activates in the 2nd time of setting test reagent
Agent, 10-40mg/L hydrogenated phospholipid, 10-50mmol/L CaCl2With 20-80mmol/L pH=7-8 buffer solution.
3. lupus anticoagulant analyte detection kit as claimed in claim 1, it is characterised in that the 1st time of setting test
Clotting factor activator in reagent and the 2nd time of setting test reagent is RVV-X, buffer solution be trishydroxymethylaminomethane-
Hydrochloric acid solution.
4. lupus anticoagulant analyte detection kit as claimed in claim 1, it is characterised in that the 1st time of setting test
Also protected in reagent and the 2nd time of setting test reagent comprising protein protective agent, preservative, ionic strength adjustor and clotting factor
Protect agent.
5. lupus anticoagulant analyte detection kit as claimed in claim 4, it is characterised in that the 1st time of setting test
Also respectively comprising 22-70g/L protein protective agents, 0.03-0.05% preservatives, 80- in reagent and the 2nd time of setting test reagent
300mmol/L ionic strength adjustors and 1-6g/L clotting factor protective agents, above-mentioned percentage are percentage by volume.
6. the lupus anticoagulant analyte detection kit as described in any one of claim 4 or 5, it is characterised in that the albumen is protected
Protect the compound that agent is component A and B component, the component A be BSA, casein or skimmed milk power, the B component be trehalose,
Mannitol or sucrose；Described preservative is Proclin300, and described ionic strength adjustor is NaCl, KCl or MgCl2,
Described clotting factor protective agent is glycine, histidine or arginine.
7. the determination methods of lupus anticoagulant presence or absence, using the kit described in claim any one of 1-6, its feature exists
In comprising the following steps：
S1, >=30 people's health normal plasmas are mixed with the 1st time of setting test reagent, the 2nd time of setting test reagent respectively,
Obtain the setting time scope T1 of normal plasma the 1st, the setting time of normal plasma the 1st and the 2nd setting time ratio range T2；
S2, the test plasma gathered from subject is divided into two parts；
S3, one of will be divided into two parts in S2 and the 1st time of setting test reagent mixing determines the 1st setting time；
S4, according to the 1st setting time and T1 relation, judge whether testing sample contains lupus anticoagulant.
8. the determination methods of lupus anticoagulant presence or absence as claimed in claim 7, it is characterised in that after the S4 steps also
Including：
S5, other a and the 2nd time of setting test reagent being divided into two parts in S2 is mixed to determine the 2nd setting time；
S6, judge whether above-mentioned test plasma contains lupus anticoagulant with the ratio of the 1st setting time and the 2nd setting time.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710355548.1A CN107589251B (en) | 2017-05-19 | 2017-05-19 | Reagent kit for detecting lupus anticoagulant and method for determining presence or absence of lupus anticoagulant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710355548.1A CN107589251B (en) | 2017-05-19 | 2017-05-19 | Reagent kit for detecting lupus anticoagulant and method for determining presence or absence of lupus anticoagulant |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107589251A true CN107589251A (en) | 2018-01-16 |
| CN107589251B CN107589251B (en) | 2020-04-28 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109211729A (en) * | 2018-10-29 | 2019-01-15 | 杨忠思 | A kind of blood stagnation condition checkout gear and method for precisely medical treatment detection |
| CN109521204A (en) * | 2018-12-07 | 2019-03-26 | 中国人民解放军陆军军医大学第附属医院 | A kind of lupus anticoagulant detection reagent and preparation method thereof and application method |
| CN110133304A (en) * | 2019-05-21 | 2019-08-16 | 北京赛科希德科技股份有限公司 | Composition, the reagent containing the composition and its application |
| CN111707837A (en) * | 2020-05-29 | 2020-09-25 | 上海太阳生物技术有限公司 | Lupus anticoagulant confirmation kit (coagulation method) |
| CN112433057A (en) * | 2020-11-10 | 2021-03-02 | 北京美创新跃医疗器械有限公司 | Screening reagent for lupus anticoagulant and preparation method thereof |
| CN112557665A (en) * | 2020-11-10 | 2021-03-26 | 北京美创新跃医疗器械有限公司 | Confirmation reagent for lupus anticoagulant and preparation method thereof |
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| CN102384979A (en) * | 2010-08-26 | 2012-03-21 | 希森美康株式会社 | Reagent kit for detecting lupus anticoagulant and method of determining presence or absence of lupus anticoagulant |
| JP2013205087A (en) * | 2012-03-27 | 2013-10-07 | Sysmex Corp | Reagent for coagulation time measurement, reagent kit for coagulation time measurement and coagulation time measuring method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109211729A (en) * | 2018-10-29 | 2019-01-15 | 杨忠思 | A kind of blood stagnation condition checkout gear and method for precisely medical treatment detection |
| CN109521204A (en) * | 2018-12-07 | 2019-03-26 | 中国人民解放军陆军军医大学第附属医院 | A kind of lupus anticoagulant detection reagent and preparation method thereof and application method |
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| CN111707837A (en) * | 2020-05-29 | 2020-09-25 | 上海太阳生物技术有限公司 | Lupus anticoagulant confirmation kit (coagulation method) |
| CN112433057A (en) * | 2020-11-10 | 2021-03-02 | 北京美创新跃医疗器械有限公司 | Screening reagent for lupus anticoagulant and preparation method thereof |
| CN112557665A (en) * | 2020-11-10 | 2021-03-26 | 北京美创新跃医疗器械有限公司 | Confirmation reagent for lupus anticoagulant and preparation method thereof |
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