CN106468715A - A kind of cervical cancer urine detection reagent and its preparation method and application - Google Patents
A kind of cervical cancer urine detection reagent and its preparation method and application Download PDFInfo
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- CN106468715A CN106468715A CN201610452206.7A CN201610452206A CN106468715A CN 106468715 A CN106468715 A CN 106468715A CN 201610452206 A CN201610452206 A CN 201610452206A CN 106468715 A CN106468715 A CN 106468715A
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Abstract
The present invention relates to a kind of cervical cancer urine detection reagent and its preparation method and application, include solution containing zinc ion compound or the mixed liquor of mercurous ionic compound solution, phosphotungstate reagents and buffer solution, the mixed proportion of solution containing zinc ion compound or mercurous ionic compound solution, phosphotungstate reagents and buffer solution is 0.5:1:1.The concentration of solution containing zinc ion compound or mercurous ionic compound solution is 0.05mol ~ 0.5mol/L, and phosphotungstate reagents comprise the aqueous solution of following component content：Sodium tungstate 40g ~ 58g/380ml ~ 650ml, strong phosphoric acid 85%/30ml ~ 50ml, concentrated hydrochloric acid 20ml ~ 50ml, lithium sulfate 30g ~ 55g, buffer solution is acetic acid/sodium acetate buffer solution, and its pH value is PH5 ~ PH6.The cervical cancer urine detection reagent of present invention preparation passes through to detect the content of the sulfydryl metabolite in urine, determine whether that cervical cancer is positive, it is not required to the instrument and equipment assisting, do not need cold preservation, establish a kind of quick, easy, sensitive, special cervical cancer detectable, to reaching the purpose to cervical cancer early screening.
Description
Technical field
The present invention relates to external diagnosis reagent technical field is and in particular to a kind of cervical cancer urine detection reagent and its preparation
Methods and applications.
Background technology
Cervical cancer is one of common malignant tumor of women worldwide, in the malignant tumor of harm women's health, cervix uteri
Cancer is listed in the 3rd.According to statistics, in all new cancer of global women, the ratio shared by cervical cancer of newly sending out is within 2008
9%, it is only second to breast carcinoma and colorectal carcinoma.In China, newly send out Patients with Cervical Cancer about 150,000 according to statistics every year.Already public
Recognize Detection of high risk human papillomavirus（HR-HPV）Persistent infection be cervix uteri carcinogenesis essential condition.But infect women from HR-HPV
Induction Cervical intraepitheliaI neoplasia（CIN）Develop into human invasive cervix neoplasms（ICC）About need the 5-20 year, this is to find by examination
And the M ＆ M treating precancerous lesion to reduce cervical cancer has won the time.
Since nearest 50 years, due to the application of effective screening method and strategy, the examination popularity rate of cervical cancer is able to relatively
It is greatly improved, worldwide, the mortality rate of cervical cancer has and significantly declines, especially in some flourishing states
Family.In our countries, although for the whole country, the mortality rate of cervical cancer has decline by a relatively large margin, this ratio
Example distribution unbalanced, the central and west regions of such as China, the population being engaged in agriculture account for the very big ratio of total population, in the middle part of it
Region-by-region is fallen behind due to economic condition, and scientific knowledge popularization face is narrow, and health resourceses lack, and people's health consciousness is poor, and this is just
These regional cervical cancer pathogenesis rates and mortality rate is led to maintain higher level all the time.In recent years, suffer from the number of cervical cancer
Assume the trend increasing year by year, ill women is also increasingly younger, causes global great attention.World Health Organization (WHO)
（WHO）Point out in coming 10 years, take higher and effective measures derepress, cervical cancer develops, otherwise, die from
The number of cervical cancer will rise 25%.
With scientific and technical development, breakthrough progress is achieved to the Study of Etiology of cervical cancer, compares one at present
Cause conclusion be：The Etiological of cervical cancer is human papillomavirus (HPV) infection.Hereafter again through epidemiology and biology
Substantial amounts of study and improve it was therefore concluded that：Continue high-risk HPV infection be cause necessity of cervical cancer and Cervical intraepitheliaI neoplasia because
Element.Cervical cancer pathogenesis are slower, and its generation, development are one and gradually accumulate, by being gradient to mutation, by the mistake of quantitative change to qualitative change
Journey.Continuous by the initial high-risk HPV infection of persistence to finally developing into this process of cervical cancer, it often to experience with
Next stage：Continue high-risk HPV infection → cervical dysplasia and carcinoma in situs of cervix (CIN I, CIN II and CIN III)
→ micro-infiltrating carcinoma → infiltrating carcinoma.Confirm that developing into invasive carcinoma of cervix (CIS) from high-risk HPV infection takes around through research
7 12 years, there is a longer precancerous lesion phase during this, be reversible.Its clear and definite paathogenic factor is eased up
Slow evolution be numerous gynaecologists timely and effectively intervene cervical cancer occurrence and development provide preciousness opportunity.And
Substantial amounts of clinical practice shows, can find cervical precancerous change early by taking cost-effective screening method, then
Take positive measure go treat, for help numerous women away from cervical cancer threat, reduce cervical cancer sickness rate and
Mortality rate has immeasurable meaning.
The malignant tumor worldwide carrying out examination of the unique suggestion of World Health Organization (WHO) is cervical cancer.Cervical cancer
Prevention target in the world is：On the one hand so that the number of women participating in cervical carcinoma screening is continuously increased, progressively expand
The scope of adult greatly woman uterus cancer examination, particularly nowadays cervical cancer patient tends to rejuvenation, and young patient more should enjoy
Concern；On the other hand, provide accurate screening method to every women suffering from cervical cancer as far as possible it is ensured that screening method itself
There is no problem for quality and effectiveness.For its prevention, new direction and prospect, mesh are provided for etiologic etiological breakthrough of cervical cancer
Front HPV-DNA detection is clinically progressively promoted as a kind of main screening method of cervical cancer；2006, HPV's was preventative
Vaccine also successfully lists.But, relatively costly due to vaccine, and whether it has preventive effect in crowd and still needs
The time of 20-30 is verified, for the numerous women of developing country, realizes vaccine prevention at present less
Reality.Therefore, the needs of developing country's national conditions are complied with, within current and a very long time in future, the examination of cervical cancer is still
It is so a kind of important means of cervical cancer preventing and treating.So, according to the practical situation of China, clinically how to select one kind both accurate
Truly have effect and economic and practical, and the screening methods of cervical cancer easily being accepted by women is particularly important.
Clinically the screening method of cervical cancer mainly has observation method of naked eye, cytolgical examination method and HPV detection method at present.Carefully
Born of the same parents' inspection is quick, inexpensive because of it, is current most common method, still plays an important role in cervix uteri examination, and many
Country is as conventional sieve project.But cytolgical examination is not very reliable, there is higher false negative, it usually needs repeat to examine
Look into the sensitivity that could obtain satisfaction, and sensitivity in adenocarcinoma detection for the Pap smear is lower than scale cancer.Although thin layer liquid-based
The appearance of the new techniques such as cytology improves cytological film-making problem, but does not thoroughly change the low present situation of its sensitivity,
This also governs the development of cytology's examination.HPV detection is another important method of cervical carcinoma screening.In European countries, HPV
Detection is replacing cytolgical examination, becomes the first-selection of cervical lesionses primary dcreening operation.HPV detection has very high sensitivity, in full-page proof
High risk group can be found, than cytology examination more effectively in this sieve.But the infection of HPV is very universal, according to statistics, property is lived
Jump women's lifelong accumulation infection rate may be up to 80%.This infection is typically transient, can be clear in time by self immune system
Remove, do not lead to the generation of height CIN and cervical cancer.Thus, HPV detection generally has higher false positive, especially in youth
In women, its specificity is lower.
Therefore, from the angle of cervical cancer sieve and early diagnosiss it is necessary to find new means to substitute or to assist
Routine screening method carries out comprehensive descision, makes the examination of cervical cancer more accurately and reliable.Using urine screening cervical cancer, overcome
The deficiency of conventional inspecting method, on the one hand enhances the compliance of experimenter, makes to be added to the women of cervical carcinoma screening significantly
Increase；On the other hand, for some non-sexual intercourse or be reluctant that the women carrying out gynecologial examination provides new cervical cancer sieve
The approach looked into, these advantages can improve the cervical carcinoma screening rate of China, promote the prevention of China's cervical cancer and control work effect
Rate, has urgent realistic meaning for the sickness rate reducing cervical cancer and its mortality rate of reduction.
Although existing cervical cancer urine detection test kit is a kind of fast and convenient method, test kit yet suffers from
The reagent holding time in composition solution type is more, structure is more complicated, operating procedure is many etc. defect, and mentioned reagent box is short,
Stability is poor.Prior art needs to be further improved and enhanced.
Content of the invention
The technical problem to be solved is, provide a kind of cervical cancer urine detection reagent and preparation method thereof and
Application, by detecting the content of sulfydryl metabolite in Urine in Patients, judges whether patient is that cervical cancer is positive, is not required to the instrument assisting
Device equipment, it is not necessary to cold preservation, establishes a kind of quick, easy, sensitive, special cervical cancer detectable, to reaching to palace
The purpose of neck cancer early screening.
The present inventor the cervix uteri carcinoma marker in urine is studied it has unexpectedly been found that, sulfhydryl compound is in Healthy People
Urine in content very low, and content significantly raises in the urine of cervical cancer patient.In order to verify this mark and palace
Specific involvement between neck cancer, the present inventor further study sulfhydryl compound in other types cancer patient's urine
Content, has been surprisingly found that and there is no the situation occurring that this protein content raises simultaneously.Therefore, sulfhydryl compound can conduct
The diagnosis sign thing of cervical cancer in urine.Technical scheme is exactly to be made with sulfhydryl compound present in urine
To detect that being present in of cervical cancer is no for target.
The present invention is achieved in that a kind of cervical cancer urine detection reagent of offer, includes molten containing zinc ion compound
Liquid or the mixed liquor of mercurous ionic compound solution, phosphotungstate reagents and buffer solution, described molten containing zinc ion compound
The mixed proportion of liquid or mercurous ionic compound solution, phosphotungstate reagents and buffer solution is 0.5:1:1；Described containing zinc from
The concentration of sub- compound solution or mercurous ionic compound solution is 0.05mol ~ 0.5mol/L, and described phosphotungstate reagents comprise
The aqueous solution of following component content：Sodium tungstate 40g ~ 58g/380ml ~ 650ml, strong phosphoric acid 85%/30ml ~ 50ml, concentrated hydrochloric acid 20ml
~ 50ml, lithium sulfate 30g ~ 55g, described buffer solution is acetic acid/sodium acetate buffer solution, and the pH value of described buffer solution is PH5
~PH6.
Further, described solution containing zinc ion compound is liquor zinci chloridi, and described mercurous ionic compound solution is
Mercuric chloride solution.
Further, the concentration of described mercuric chloride solution is 0.05mol/L；Described sodium tungstate 45g/380ml, strong phosphoric acid
85%/30ml, concentrated hydrochloric acid 30ml, lithium sulfate 30g, finally plus distilled water is settled to 1000ml；Described sodium acetate 82.3g/
1000ml, addition acetic acid adjust pH value to the acetic acid/sodium acetate buffer solution of PH5.3 ~ PH5.5.
Further, the concentration of described mercuric chloride solution is 0.08mol/L；Described sodium tungstate 51g/400ml, strong phosphoric acid
85%/37ml, concentrated hydrochloric acid 20ml, lithium sulfate 38g, finally plus distilled water is settled to 1000ml；Described sodium acetate 70g/1000ml,
Acetic acid is added to adjust pH value to the acetic acid/sodium acetate buffer solution of PH5.5 ~ PH5.7.
Further, the concentration of described liquor zinci chloridi is 0.1mol/L；Described sodium tungstate 58g/520ml, strong phosphoric acid
85%/40ml, concentrated hydrochloric acid 30ml, lithium sulfate 40g, finally plus distilled water is settled to 1000ml；Described sodium acetate 65g/1000ml,
Acetic acid is added to adjust pH value to the acetic acid/sodium acetate buffer solution of PH5 ~ PH5.3.
Further, the concentration of described liquor zinci chloridi is 0.5mol/L；Described sodium tungstate 40g/650ml, strong phosphoric acid
85%/50ml, concentrated hydrochloric acid 50ml, lithium sulfate 55g, finally plus distilled water is settled to 1000ml；Described sodium acetate 95g/1000ml,
Acetic acid is added to adjust pH value to the acetic acid/sodium acetate buffer solution of PH5.7 ~ PH6.
The present invention also provides a kind of preparation method of cervical cancer urine detection reagent as the aforementioned, including：
（1）Solution containing zinc ion compound or the preparation of mercurous ionic compound solution：By concentration 0.05mol ~ 0.5mol/L
Requirement weigh be dissolved in containing zinc ion compound or mercurous ionic compound standby after distilled water；
（2）The preparation of phosphotungstate reagents：Weigh sodium tungstate 40g ~ 58g, be dissolved in distilled water 500ml ~ 650ml, add 85% dense phosphorus
Sour 30ml ~ 50ml, adds concentrated hydrochloric acid 30ml ~ 50ml, flows back 2 hours ~ 4 hours, is settled to 1000ml, is eventually adding sulfur after cooling
Sour lithium 30g ~ 55g, saves backup；
（3）The preparation of buffer solution：65g ~ 82.3g sodium acetate is taken to dissolve in the water of 1000ml, with acetic acid tune pH value to PH 5
To PH6；
（4）Phosphotungstate reagents are mixed with buffer solution：By（2）Step and the（3）The phosphotungstate reagents of step preparation are molten with buffering
Liquid presses 1:1 ratio mix homogeneously, obtains phosphotungstic acid buffer solution；
（5）By（1）Step and the（4）Preparation solution containing zinc ion compound or mercurous ionic compound solution 0.5ml and
Phosphotungstic acid buffer solution 2ml mix homogeneously makes the mixed liquor of 2.5ml, by the ampoule bottle of this mixed liquor sterile packaged to 10ml
In.
The invention also discloses carry out examination and the auxiliary diagnosis of cervical cancer using aforesaid cervical cancer urine detection reagent
The application of aspect.
The method of the examination and auxiliary diagnosis that carry out cervical cancer using the cervical cancer urine detection reagent of the present invention is：
Take women patient urina sanguinis 0.5ml, add in the reagent bottle of cervical cancer urine detection reagent filling the present invention, shake up standing
5 ~ 10 minutes, the such as solution colour in reagent bottle was shown as colourless or milky, then result is feminine gender, illustrates that patient does not suffer from palace
Neck cancer.As the solution colour in reagent bottle is shown as blue, then result is the positive, illustrates that patient suffers from cervical cancer.
Compared with prior art, the beneficial effect of cervical cancer urine detection reagent of the present invention and its preparation method and application
It is：
（1）By the use of patient urine as detection sample, patient's no pain；
（2）Preparation is convenient, low cost；
（3）Detection method is simple to operate, and specificity is high, and diagnostic result is accurate；
（4）The early stage of the extensive examination and High-risk Population of Cervical Carcinoma that can be used for cervical cancer takes precautions against.
Specific embodiment
In order that the technical problem to be solved, technical scheme and beneficial effect become more apparent, below tie
Close embodiment, the present invention will be described in further detail.It should be appreciated that specific embodiment described herein is only in order to solve
Release the present invention, be not intended to limit the present invention.And, in the following example, the experimental technique of unreceipted actual conditions, generally presses
Conventional meanses according to this area are carried out.
Embodiment 1：Preparation cervical cancer urine detection reagent 1000
（1）Prepared by mercuric chloride solution：Weigh mercuric chloride by the requirement of concentration 0.05mol/L and be dissolved in after distilled water and be settled to 500ml
Standby.
（2）The preparation of phosphotungstate reagents：Weigh sodium tungstate 45g, be dissolved in distilled water 380ml, add 85% strong phosphoric acid
30ml, adds concentrated hydrochloric acid 30-50ml, and backflow is boiled 2 hours, adds lithium sulfate 30g, finally plus distilled water is settled to after cooling
1000ml saves backup.
（3）The preparation of buffer：Sodium acetate 82.3g/1000ml, addition acetic acid adjust pH value to PH5.3 ~ PH5.5, preserve standby
With.
（4）Phosphotungstate reagents are mixed with buffer solution：By（2）Step and the（3）Walk the phosphotungstate reagents of preparation and delay
Rush solution by 1:1 ratio mix homogeneously, obtains phosphotungstic acid buffer solution.
（5）By（1）Step and the（4）The mercuric chloride solution 0.5ml of preparation and phosphotungstic acid buffer solution 2ml mix homogeneously system
Become the mixed liquor of 2.5ml, by this mixed liquor sterile packaged in the ampoule bottle of 10ml, obtain a cervical cancer urine detection examination
Agent.
（6）Repeat（5）It is sequentially completed the preparation of 1000 cervical cancer urine detection reagent.
Embodiment 2：Another kind of cervical cancer urine detection reagent 1000 of preparation
（1）Prepared by mercuric chloride solution：Weigh mercuric chloride by the requirement of concentration 0.08mol/L and be dissolved in after distilled water and be settled to 500ml
Standby.
（2）The preparation of phosphotungstate reagents：Weigh sodium tungstate 51g, be dissolved in distilled water 400ml, add 85% strong phosphoric acid
37ml, adds concentrated hydrochloric acid 20ml, and backflow is boiled 2 hours, adds lithium sulfate 38g dissolving, finally plus distilled water is settled to after cooling
1000ml, saves backup.
（3）The preparation of buffer：Sodium acetate 70g/1000ml, addition acetic acid adjust pH value to PH5.5 ~ PH5.7, preserve standby
With.
（4）Phosphotungstate reagents are mixed with buffer solution：By（2）Step and the（3）Walk the phosphotungstate reagents of preparation and delay
Rush solution by 1:1 ratio mix homogeneously, obtains phosphotungstic acid buffer solution.
（5）By（1）Step and the（4）The mercuric chloride solution 0.5ml of preparation and phosphotungstic acid buffer solution 2ml mix homogeneously system
Become the mixed liquor of 2.5ml, by this mixed liquor sterile packaged in the ampoule bottle of 10ml, obtain a cervical cancer urine detection examination
Agent.
（6）Repeat（5）It is sequentially completed the preparation of 1000 cervical cancer urine detection reagent.
Embodiment 3：Prepare another cervical cancer urine detection reagent 1000
（1）Prepared by liquor zinci chloridi：Weigh zinc chloride by the requirement of concentration 0.1mol/L and be dissolved in after distilled water and be settled to 500ml
Standby.
（2）The preparation of phosphotungstate reagents：Weigh sodium tungstate 58g, be dissolved in distilled water 520ml, add 85% strong phosphoric acid
40ml, adds concentrated hydrochloric acid 30ml, and backflow is boiled 2 hours, adds lithium sulfate 40g dissolving, finally plus distilled water is settled to after cooling
1000ml, saves backup.
（3）The preparation of buffer：Sodium acetate 65g/1000ml, addition acetic acid adjust pH value to PH5 ~ PH5.3, save backup.
（4）Phosphotungstate reagents are mixed with buffer solution：By（2）Step and the（3）Walk the phosphotungstate reagents of preparation and delay
Rush solution by 1:1 ratio mix homogeneously, obtains phosphotungstic acid buffer solution.
（5）By（1）Step and the（4）The liquor zinci chloridi 0.5ml of preparation and phosphotungstic acid buffer solution 2ml mix homogeneously system
Become the mixed liquor of 2.5ml, by this mixed liquor sterile packaged in the ampoule bottle of 10ml, obtain a cervical cancer urine detection examination
Agent.
（6）Repeat（5）It is sequentially completed the preparation of 1000 cervical cancer urine detection reagent.
Embodiment 4：Prepare another cervical cancer urine detection reagent 1000
（1）Prepared by liquor zinci chloridi：Weigh zinc chloride by the requirement of concentration 0.5mol/L and be dissolved in after distilled water and be settled to 500ml
Standby.
（2）The preparation of phosphotungstate reagents：Weigh sodium tungstate 40g, be dissolved in distilled water 650ml, add 85% strong phosphoric acid
50ml, adds concentrated hydrochloric acid 50ml, and backflow is boiled 2 hours, adds lithium sulfate 55g dissolving, finally plus distilled water is settled to after cooling
1000ml, saves backup.
（3）The preparation of buffer：Sodium acetate 95g/1000ml, addition acetic acid adjust pH value to PH5.7 ~ PH6, save backup.
（4）Phosphotungstate reagents are mixed with buffer solution：By（2）Step and the（3）Walk the phosphotungstate reagents of preparation and delay
Rush solution by 1:1 ratio mix homogeneously, obtains phosphotungstic acid buffer solution.
（5）By（1）Step and the（4）The liquor zinci chloridi 0.5ml of preparation and phosphotungstic acid buffer solution 2ml mix homogeneously system
Become the mixed liquor of 2.5ml, by this mixed liquor sterile packaged in the ampoule bottle of 10ml, obtain a cervical cancer urine detection examination
Agent.
（6）Repeat（5）It is sequentially completed the preparation of 1000 cervical cancer urine detection reagent.
Embodiment 5：The cervix uteri urine cancer detectable of the present invention and the comparison of other cervical cancer detection methods
1. subjects：
Choose OPD of Obs ＆ Gyn go to a doctor patient's totally 378 female subjects, age 25-52 year.Inclusion criteria：Sexual intercourse history,
Non-pregnancy, age of sucking and menstrual phase, no malignant tumor history, ametria excises medical history, no operation on cervix treatment history.All tested
Person all signs Informed Consent Form.
2. detection method：
（1）The cervical cancer detectable of the present invention is prepared according to the step of above-described embodiment 1.
（2）Liquid-based cytology detection method：Wipe cervical surface mucus, cervical canal is stretched into using special cervix cellss brush
At interior 1 cm, rotation was taken out after 4 ~ 6 weeks in the direction of the clock, collected the cell that cervix cellss paint, and is stored in and preserves equipped with cell
In the special bottle of liquid, by preserving the cell in liquid, the Pap smear being evenly distributed is made using natural sedimentation method, divided using TBS
Level diagnostic criteria diagosis method carries out diagosis.Cervical cancer is determine whether according to liquid basal cell inspection result positive.
（3）Pathological biopsy：Cervix uteri multiple punch biopsy is carried out for all experimenters or neck tube is scratched and scraped, take tissue to do pathology inspection
Look into, whether diagnosis suffers from cervical cancer.
3. collection data is processed：
All experimenter is detected using the cervical cancer urine detection reagent of the present invention respectively simultaneously, obtains testing result 1,
Detected using liquid-based cytology detection method, obtained testing result 2, using pathological biopsy detection, obtained testing result 3,
And testing result is compared, calculate the ginseng such as sensitivity, specificity, accuracy, positive predictive value, negative predictive value more simultaneously
Numerical value is compared.Wherein, sensitivity=true positives/(true positives+false negative), specificity=true negative/(false positive+Kidney-Yin
Property), accuracy=(true positives+true negative)/total number of persons, positive predictive value=true positives/(true positives+false positive), negative prediction
Value=true negative/(false negative+true negative).Cervical cancer（+）It is denoted as cervical cancer tests positive, cervical cancer（-）It is denoted as palace
Neck cancer detection is negative.
Have 378 experimenters and participate in this examination, the testing result that three kinds of detection methods obtain is shown in table 1, detection
The sensitivity of result, specificity, accuracy, positive predictive value, negative predictive value are shown in table 2.
The different detection method of table 1. and the comparison of pathological examination results
The comparison of the different testing result indices of table 2.
In combination with Tables 1 and 2 as can be seen that the present invention detects Urinary Specific mark come diagnosing cervical by sulfydryl
Every Testing index of method will be better than liquid-based cytology detection method, the urine detection reagent specificity of the present invention is described
By force, accuracy rate is high, can be used as effective cervical carcinoma screening reagent.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all essences in the present invention
Any modification, equivalent and improvement made within god and principle etc., should be included within the scope of the present invention.
Claims (8)
1. a kind of cervical cancer urine detection reagent is it is characterised in that including solution containing zinc ion compound or containing mercury ion
The mixed liquor of compound solution, phosphotungstate reagents and buffer solution, described solution containing zinc ion compound or contain mercury ion
The mixed proportion of compound solution, phosphotungstate reagents and buffer solution is 0.5:1:1；Described solution containing zinc ion compound or
The concentration of the mercurous ionic compound solution of person is 0.05mol ~ 0.5mol/L, and described phosphotungstate reagents comprise following component content
Aqueous solution：Sodium tungstate 40g ~ 58g/380ml ~ 650ml, strong phosphoric acid 85%/30ml ~ 50ml, concentrated hydrochloric acid 20ml ~ 50ml, lithium sulfate
30g ~ 55g, described buffer solution is acetic acid/sodium acetate buffer solution, and the pH value of described buffer solution is PH5 ~ PH6.
2. cervical cancer urine detection reagent as claimed in claim 1 is it is characterised in that described solution containing zinc ion compound is
Liquor zinci chloridi, described mercurous ionic compound solution is mercuric chloride solution.
3. cervical cancer urine detection reagent as claimed in claim 2 is it is characterised in that the concentration of described mercuric chloride solution is
0.05mol/L；Described sodium tungstate 45g/380ml, strong phosphoric acid 85%/30ml, concentrated hydrochloric acid 30ml, lithium sulfate 30g, finally plus distillation
Water is settled to 1000ml；Described sodium acetate 82.3g/1000ml, addition acetic acid adjust pH value to the acetic acid/acetic acid of PH5.3 ~ PH5.5
Sodium buffer solution.
4. cervical cancer urine detection reagent as claimed in claim 2 is it is characterised in that the concentration of described mercuric chloride solution is
0.08mol/L；Described sodium tungstate 51g/400ml, strong phosphoric acid 85%/37ml, concentrated hydrochloric acid 20ml, lithium sulfate 38g, finally plus distillation
Water is settled to 1000ml；Described sodium acetate 70g/1000ml, addition acetic acid adjust pH value to the acetic acid/sodium acetate of PH5.5 ~ PH5.7
Buffer solution.
5. cervical cancer urine detection reagent as claimed in claim 2 is it is characterised in that the concentration of described liquor zinci chloridi is
0.1mol/L；Described sodium tungstate 58g/520ml, strong phosphoric acid 85%/40ml, concentrated hydrochloric acid 30ml, lithium sulfate 40g, finally plus distilled water
It is settled to 1000ml；Described sodium acetate 65g/1000ml, addition acetic acid adjust pH value to the acetic acid/acetate buffer of PH5 ~ PH5.3
Solution.
6. cervical cancer urine detection reagent as claimed in claim 2 is it is characterised in that the concentration of described liquor zinci chloridi is
0.5mol/L；Described sodium tungstate 40g/650ml, strong phosphoric acid 85%/50ml, concentrated hydrochloric acid 50ml, lithium sulfate 55g, finally plus distilled water
It is settled to 1000ml；Described sodium acetate 95g/1000ml, addition acetic acid adjust pH value to the acetic acid/acetate buffer of PH5.7 ~ PH6
Solution.
7. a kind of preparation method of cervical cancer urine detection reagent as claimed in claim 1 or 2 is it is characterised in that include：
（1）Solution containing zinc ion compound or the preparation of mercurous ionic compound solution：By concentration 0.05mol ~ 0.5mol/L
Requirement weigh be dissolved in containing zinc ion compound or mercurous ionic compound standby after distilled water；
（2）The preparation of phosphotungstate reagents：Weigh sodium tungstate 40g ~ 58g, be dissolved in distilled water 500ml ~ 650ml, add 85% dense phosphorus
Sour 30ml ~ 50ml, adds concentrated hydrochloric acid 30ml ~ 50ml, flows back 2 hours ~ 4 hours, is settled to 1000ml, is eventually adding sulfur after cooling
Sour lithium 30g ~ 55g, saves backup；
（3）The preparation of buffer solution：65g ~ 82.3g sodium acetate is taken to dissolve in the water of 1000ml, with acetic acid tune pH value to PH 5
To PH6；
（4）Phosphotungstate reagents are mixed with buffer solution：By（2）Step and the（3）The phosphotungstate reagents of step preparation are molten with buffering
Liquid presses 1:1 ratio mix homogeneously, obtains phosphotungstic acid buffer solution；
（5）By（1）Step and the（4）Preparation solution containing zinc ion compound or mercurous ionic compound solution 0.5ml and
Phosphotungstic acid buffer solution 2ml mix homogeneously makes the mixed liquor of 2.5ml, by the ampoule bottle of this mixed liquor sterile packaged to 10ml
In.
8. the application of the cervical cancer urine detection reagent as described in any one in claim 1 to 6 is it is characterised in that described
Cervical cancer urine detection reagent is applied to examination and the auxiliary diagnosis aspect of cervical cancer.
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