CN109971847A - Application of the Calponin-h2 as the marker of Diagnosis of Ectopic Pregnancy - Google Patents
Application of the Calponin-h2 as the marker of Diagnosis of Ectopic Pregnancy Download PDFInfo
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Abstract
Application the present invention relates to Calponin-h2 (CNN2) as the marker of Diagnosis of Ectopic Pregnancy.The invention firstly discloses the specific serum markers that CNN2 can be used as ectopic pregnancy early diagnosis, have good sensitivity and specificity.The auxiliary diagnosis that ectopic pregnancy is carried out using CNN2 can promote the timeliness and accuracy of diagnosis, effectively improve the early diagnostic rate of ectopic pregnancy.
Description
Technical field
The invention belongs to diagnostic flag fields, more particularly it relates to ectopic pregnancy early diagnoses, with
Application of the Calponin-h2 as the marker of Diagnosis of Ectopic Pregnancy.
Background technique
Ectopic pregnancy refers to fertilized eggs position implantation other than cavity of uterine body, is one of most common acute abdomen of gynemetrics.
Rupture of tubal pregnancy can cause the complication such as intraperitoneal hemorrhage, infection, shock, and serious person can jeopardize patient vitals.Ectopic pregnancy
It is one of most common acute abdomen of gynemetrics, can lead to a large amount of intraperitoneal hemorrhages, patient's shock is even dead.In developed country, 9
~13% maternal death is caused by ectopic pregnancy, and in developing country, this ratio up to 10~30%.
The method of early diagnosis of ectopic pregnancy at present is not perfect.Due to lacking the blood serum designated object of specificity, clinically
It relies primarily on transvaginal sonography to be diagnosed, or is diagnosed until symptom is presented, be extremely easy to that the delay in diagnosis occurs,
And lead to serious consequence.Although some diagnostic works can be carried out by transvaginal sonography, however, there is about 8~31% patient
Result of ultrasonography is difficult to clear and further operation or laboratory is needed to check to make a definite diagnosis.The early diagnosis of ectopic pregnancy is to work as
Difficult point and pain spot in preceding clinical position.
Although recent 20 years researcher is dedicated to finding the specific serum marker of ectopic pregnancy, since dystopy is pregnant
The etiology unknown being pregnent, while effective animal model can not be established, thus correlative study be in progress so far it is little.It has now been found that
Some potential markers for being likely to become early diagnosis ectopic pregnancy include vascular endothelial growth factor (Wang, D.J., et
al.,Association study of vascular endothelial growth factor gene
polymorphisms with ectopic pregnancy in Chinese women.Clin Exp Obstet
Gynecol, 2014.41 (6): p.665-70), pregnant associated serum albumen-A (Rausch, M.E.and K.T.Barnhart,
Serum biomarkers for detecting ectopic pregnancy.Clin Obstet Gynecol,2012.55
(2): p.418-23), inhibin A (Senapati, S., et al., Predicting first trimester pregnancy
outcome:derivation of a multiple marker test.Fertil Steril,2016.106(7):
P.1725-1732e3), activin A (Wijayarathna, R.and D.M.de Kretser, Activins in
Reproductive biology and beyond.Hum Reprod Update, 2016.22 (3)), miR-323 etc., but this
The sensitivity of a little markers and specificity be not high, and research conclusion is also inconsistent, and most researchs are only retrospective analysis or disease
Example check analysis, lacks prospective cohort study, thus still have it is more it is insufficient (Senapati, S.and K.T.Barnhart,
Biomarkers for ectopic pregnancy and pregnancy of unknown location.Fertil
Steril,2013.99(4):p.1107-16).Table 1 has carried out ectopic pregnancy blood serum designated object reported at present simple total
Knot.
Table 1, ectopic pregnancy blood serum designated object
Blood β-hCG is the most common serum markers of current diagnosis ectopic pregnancy.Single blood β-hCG detection is only capable of prompting pregnant
It is pregnent and cannot reflect pregnant position；Continuous monitoring blood β-hCG is then that raising has more than 53% as judgement using in β-hCG value 48h
Vigor uterine pregnancy (viable intrauterine pregnancy, vIUP) standard (American College of,
O.and Gynecologists,ACOG Practice Bulletin No.94:Medical management of
Ectopic pregnancy.Obstet Gynecol, 2008.111 (6): p.1479-85), therefore being more living as gestation
The exclusion index of power rather than the diagnosis index of ectopic pregnancy.Continuous monitoring blood β-hCG Diagnosis of Ectopic Pregnancy false sun with higher
Property and false negative rate, and it is difficult to antidiastole spontaneous abortion and ectopic pregnancy；Meanwhile patient monitors the phase in multiple blood β-hCG
Between have occur ruptured ectopic pregnancy bleeding risk.
To sum up, this field needs to find highly sensitive, the ectopic pregnancy specific serum marker of high specificity.
Summary of the invention
Application the purpose of the present invention is to provide Calponin-h2 as the marker of Diagnosis of Ectopic Pregnancy.
In the first aspect of the present invention, it is pregnant in preparation dystopy to provide Calponin-h2 (CNN2) albumen or its encoding gene
The purposes in diagnostic reagent or kit being pregnent.
In a preferred embodiment, the ectopic pregnancy are as follows: tubal pregnancy.
In another preferred example, the diagnostic reagent is selected from:
The primer of the encoding gene of specific amplification Calponin-h2 albumen；Or
The encoding gene of specific recognition Calponin-h2 albumen or the probe of its transcript；Or
Specifically bind the binding molecule of Calponin-h2 albumen；Or
The encoding gene of specific recognition Calponin-h2 albumen or the chip of its transcript.
In another preferred example, the primer of the encoding gene of the specific amplification Calponin-h2 albumen is primer
Right, nucleotide sequence is as shown in SEQ ID NO:1 and SEQ ID NO:2.
In another preferred example, the binding molecule of the specific binding Calponin-h2 albumen is antibody.
In another preferred example, the antibody is monoclonal antibody or polyclonal antibody.
In another preferred example, the diagnostic reagent is the diagnostic reagent using serum as sample to be tested, with blood plasma be to
The diagnostic reagent of sample or using peripheral blood mononuclear cells as the diagnostic reagent of sample to be tested；Preferably, be with serum be to
The diagnostic reagent of sample.
In another aspect of this invention, a kind of kit for Diagnosis of Ectopic Pregnancy is provided, is contained in the kit
Have: the diagnostic reagent of the expression or expression quantity of detection Calponin-h2 albumen or its encoding gene, or be mounted with and change diagnosis
The solid phase carrier (such as test paper or test strips) of reagent.
In a preferred embodiment, the diagnostic reagent is selected from: the encoding gene of specific amplification Calponin-h2 albumen
Primer；Specific recognition Calponin-h2 albumen encoding gene or its transcript probe；Or specific binding
The binding molecule of Calponin-h2 albumen；Specific recognition Calponin-h2 albumen encoding gene or its transcript core
Piece.
In another preferred example, the diagnostic reagent is the encoding gene of specific amplification Calponin-h2 albumen
Primer, nucleotide sequence is as shown in SEQ ID NO:1 and SEQ ID NO:2.
In another preferred example, in the kit further include: nucleic acid extraction reagent；Pcr reagent；Egg
White immunohistochemistry reagent；Enzyme chain immune response reagent；Or operation instructions.
In another aspect of this invention, a kind of method of diagnostic reagent preparing ectopic pregnancy is provided, which comprises
Using Calponin-h2 albumen or its encoding gene as object, prepare specific recognition or in conjunction with the Calponin-h2 albumen or
The reagent of its encoding gene.
In a preferred embodiment, the reagent includes:
The primer of the encoding gene of specific amplification Calponin-h2 albumen；Or
The encoding gene of specific recognition Calponin-h2 albumen or the probe of its transcript；Or
Specifically bind the binding molecule of Calponin-h2 albumen；Or
The encoding gene of specific recognition Calponin-h2 albumen or the chip of its transcript.
In another preferred example, the method also includes: by the specific recognition or in conjunction with the Calponin-h2
On the load reagents to solid phase carrier of albumen or its encoding gene；Preferably, the solid phase carrier includes but is not limited to: examination
Paper or test strips, microballoon, slide.
In another aspect of this invention, a kind of method of Diagnosis of Ectopic Pregnancy is provided, which comprises
(1) table of Calponin-h2 albumen or gene in detection subject oviduct tissue, blood plasma, serum or peripheral blood
Up to level；
(2) Calponin-h2 albumen or gene expression dose are analyzed, if the expression of conspicuousness height occurs, shows that subject is
The people at highest risk or patient of ectopic pregnancy.
In a preferred embodiment, which comprises
(1) using Calponin-h2 in mentioned-above reagent or kit detection subject's blood plasma, serum or peripheral blood
Albumen or gene expression dose；
(2) Calponin-h2 albumen or gene expression dose are analyzed, if comparison normal healthy controls crowd, Calponin-h2 egg
The expression of white or gene expression dose conspicuousness height then shows that subject is the people at highest risk or patient of ectopic pregnancy.
Other aspects of the invention are apparent to those skilled in the art due to this disclosure
's.
Detailed description of the invention
Fig. 1, PCR detect CNN2 expression in the chorionic villi of ectopic pregnancy；
Control group: uterine pregnancy chorionic villi；
Test group: ectopic pregnancy chorionic villi；
The unit of ordinate: opposite mRNA is expressed.
Fig. 2, Immunohistochemical Method detection CNN2 plant the expression of fallopian tubal part in ectopic gestation sac semina；
A, C, E are the oviduct tissue of embryonic implantation part；
B, the part oviduct tissue of D, F not far from embryonic implantation (> 2 centimetres)；
CNN1 dyes (A, B)；CNN2 dyes (C, D)；CNN3 dyes (E, F).
The ROC curve of Fig. 3, CNN2 Diagnosis of Ectopic Pregnancy.Using 56.97ng/ml as dividing value, AUC=0.955, sensitivity=
95.2%, specificity=93.7%, Youden index=0.889；Positive predictive value=96.4%, negative predictive value=
91.8%.
Fig. 4, change of serum C NN2 are lowered in the postoperative expression of ectopic pregnancy.84 patients are in the preoperative and postoperative detection change of serum C NN2 water
It is flat, and the postoperative expression downward of CNN2 (preoperative 91.55 ± 21.89, postoperative 78.99 ± 25.34), P < 0.001.
Fig. 5, change of serum C NN2 concentration and pregnant week correlation；Abscissa indicates that pregnancy number of days, ordinate indicate CNN2 concentration, often
A point represents a case-specific；
Left figure is CNN2 concentration and pregnant week relationship, r=0.024, P=0.832 in ectopic pregnancy.
Right figure is CNN2 concentration and pregnant week relationship, r=-0.004, P=0.98 in uterine pregnancy.
Specific embodiment
The present inventor passes through in-depth study, and disclosing Calponin-h2 (CNN2) for the first time can be used as ectopic pregnancy morning
The specific serum marker of phase diagnosis, has good sensitivity and specificity.The auxiliary of ectopic pregnancy is carried out using CNN2
Diagnosis can promote the timeliness and accuracy of diagnosis, effectively improve the early diagnostic rate of ectopic pregnancy.
As used in the present invention, " ectopic pregnancy " and " ectopic pregnancy " is used interchangeably, and refers to blastaea other than uterus
The disease of region implantation.
Calponin-h2
In the present invention, the amino acid sequence of term " Calponin-h2 (CNN2) albumen " and GenBank accession number
The protein sequence that AAI41834.1 is provided is substantially the same, also includes the homologous protein of CNN2 albumen.
CNN2 is a kind of actin cytoskeleton binding protein.The cell of expression CNN2 is roughly divided into three classes: (1) in life
The cell that high mechanical tension is born in the case of reason, such as the smooth muscle cell on hollow organ wall, epithelium and endothelial cell；(2) high
The cell of proliferation rate；(3) active cell is migrated.And a kind of intracellular response factors of the CNN2 as mechanical stress, by machinery
Stress is adjusted, and by interacting with cytoskeleton F- actin and a variety of vinculins, participates in the sense of cyto-mechanics signal
By with conduction, adjust cell Proliferation, differentiation, the physiological activities such as migration.The present inventor speculates that blastaea is planted in its other than uterus
When on its position such as fallopian tubal, the epithelium for planting position may cause the rapid variation of mechanical pressure.This planting process is similar
Blood platelet hemostasis: for the free floating platelet adhesion reaction in blood vessel at vascular trauma, the positioning adherency of blastaea can be in female tire
Interface forms variation that is mechanically stressed, and causing cytoskeleton tension；Then may be used when blastaea is planted in poor uterine tubal epithelium
It can cause " bigger more rapid mechanical stress ".The variation of this mechanical pressure, so that the expression of CNN2 changes.
In the embodiment of the present invention, the present inventor analyzes expression of the CNN2 in ectopic pregnancy patients.
It is detected with PCR and ImmunohistochemistryMethods Methods, discovery CNN2 expresses increase in ectopic pregnancy chorionic villi；In ectopic pregnancy embryo
High expression in the urine output tubing of bed portions point, and away from not expressing in the oviduct tissue of bed portions point or low expression.Use ELISA
Method detection finds that CNN2 is horizontal negative in non-pregnant woman's serum, and ectopic pregnancy patients change of serum C NN2 level is apparently higher than just
Normal uterine pregnancy, and 1-3 days after surgery change of serum C NN2 levels are decreased obviously.Change of serum C NN2 concentration is to menopause number of days without related simultaneously
Property.Therefore, CNN2 can be used as identification whether Ren Shen index, while again be identification ectopic pregnancy and uterine pregnancy specificity
Marker.
In the embodiment of the present invention, change of serum C NN2 is analyzed as the feasibility of Diagnosis of Ectopic Pregnancy marker.
84 ectopic pregnancies and 39 uterine pregnancy patients serum CNN2 detections are shown to determine 56.97ng/ml as boundary using ROC curve
Value, sensitivity=95.2% of CNN2 Diagnosis of Ectopic Pregnancy, specificity=93.7%.Index=0.889 Youden, positive prediction
Value is 96.4%, and negative predictive value is 91.8%, this result is substantially better than the diagnostic marker applied at present.
Diagnostic reagent and application
New discovery based on the present inventor, can be using CNN2 albumen or its encoding gene as the mark of measurement ectopic pregnancy
Object (marker).By analysis sample to be tested (sample) in the expression of CNN2 albumen or its encoding gene, thus learn by
The disease state of examination person provides foundation for the diagnosis or prognosis of disease.In a preferred embodiment, the sample to be tested or
Sample to be tested is the body fluid of patient, preferably serum.
The present invention provides specific recognition CNN2 albumen or the reagents of its encoding gene.Any recognizable CNN2 albumen or
The reagent of its encoding gene is included in the present invention, the marker as detection ectopic pregnancy.The specific recognition
The reagent of CNN2 albumen or its encoding gene is, for example: the primer of the encoding gene of specific amplification CNN2 albumen；Or specificity
Identify the encoding gene of CNN2 albumen or the probe of its transcript；Or the antibody of the anti-CNN2 albumen of specificity.
The present invention also provides the purposes of a kind of specific recognition CNN2 albumen or the reagent of its encoding gene, for detecting
Ectopic pregnancy.
The multiple technologies in this field can be used to detect the nucleic acid of CNN2 or the expression of albumen, these technologies are wrapped
Containing in the present invention.Detect the available prior art of nucleic acid such as (but not limited to): biochip technology, is gathered at probe hybridization technique
The methods of synthase chain reaction (PCR), Northern Blot.Detect albumen can using immunohistochemistry, Western Blot or
The methods of ELISA or by means of mass spectrometer etc..
As preferred embodiment of the invention, the antibody of application specific combination CNN2 albumen, with immunohistochemistry technique or
ELISA gene etc. detects the expression of CNN2 albumen.
It is well known in the art for preparing the technology of antibody.Antibody of the invention, which can be, has spy to CNN2 albumen
Anisotropic monoclonal antibody.Monoclonal antibody can use hybridoma technology to prepare (see Kohler et al., Nature 256；
495,1975；Kohler et al., Eur.J.Immunol.6:511,1976；Kohler et al., Eur.J.Immunol.6:292,
1976；Hammerling et al., In Monoclonal Antibodies and T Cell Hybridomas, Elsevier,
N.Y., 1981).The monoclonal antibody can use CNN2 albumen or protein fragments or functional areas, be obtained by immunological technique.
Further, it is also possible to be prepared using recombination method or synthesized using Peptide synthesizer.The antibody is also possible to CNN2 albumen
Polyclonal antibody with specificity.The polyclonal antibody can be prepared by conventional method, for example, can be by by institute
The CNN2 albumen stated is imported in animal and is obtained, for example, CNN2 albumen is mixed with Freund's adjuvant according to proper proportion (such as 1:1)
Animal is immunized afterwards.Animal subcutaneous injection can be used in immunization method.The animal can be selected from rabbit, sheep, mouse etc..
The reagent of the specific recognition CNN2 albumen or its encoding gene, also can be fixed in test paper, slide or its
On its solid phase carrier, it is prepared into immune colloid gold test paper or similar detection material.
As a kind of selectable mode of the invention, a kind of survey is prepared using the antibody of specific recognition CNN2 albumen
Strip comprising solid phase carrier (usually test-strips, such as test paper (item)), and be attached to described on the solid phase carrier
Antibody.For example, immune colloidal gold test strip can be prepared into.Such detection instrument is very convenient, and patient can be voluntarily
Be in and carry out the screening of ectopic pregnancy, can early detection ectopic pregnancy in time, it is easy to operate.
As another selection mode of the invention, pass through polymerase chain reaction (PCR) Fa Laifen quantitatively or semi-quantitatively
The expression and expression quantity of CNN2 gene in sample are analysed, so as to judge.Preferably, passing through real-time quantitative
Realtime-PCR realizes detection.The reagent of use is the primer of specific amplification CNN2 gene, in the nucleosides for knowing CNN2
After acid sequence, people are easy to design primer based on this.As more preferable mode of the invention, the primer is primer pair,
With nucleotide sequence shown in SEQ ID NO:1 and SEQ ID NO:2.The primer pair can expand acquisition appropriate length
Amplified production, the specificity of detection is good.
Biochip technology can also be used to carry out the detection of CNN2.After knowing the nucleotide sequence of CNN2, people
It is easy to design probe based on this.For example, ammonia is contained at 5 ' ends of probe if solid phase carrier is using modification slide or silicon wafer
Oligonucleotide probe, can be configured to solution by the poly- dT string of base modification, and then point sample instrument is by its point in modification slide or silicon wafer, row
Scheduled sequence or array are arranged into, is then fixed by standing overnight, so that it may obtain genetic chip of the invention.If few core
Thuja acid probe is without amido modified, then preparation method can also refer to existing well-known technique.
Diagnostic kit
The present invention also provides the kit for detecting ectopic pregnancy, which includes: specific recognition CNN2 egg
White or its encoding gene reagent, or it is mounted with the carrier of the reagent.The specific recognition CNN2 albumen or its coding
The reagent of gene is, for example: the primer of the encoding gene of specific amplification CNN2 albumen；Or the volume of specific recognition CNN2 albumen
The probe of code gene or its transcript；Or the antibody (monoclonal antibody or polyclonal antibody) of the anti-CNN2 albumen of specificity.
Can also contain some auxiliary reagents in the kit: nucleic acid extraction reagent (such as nucleic acid extraction liquid, phenol, chloroform,
Isoamyl alcohol, NaCl etc.)；And/or pcr reagent (such as dNTP, Taq enzyme, PCR buffer, archaeal dna polymerase)；With/
Or protein groups reagent (such as colour reagent, dimethylbenzene, ethyl alcohol, H2O2Methanol solution, antigen retrieval buffers, confining liquid, PBS etc.)；With/
Or enzyme chain immune response reagent (such as developing solution or hybridization solution).
In addition, also containing the operation instructions and/or standard operation journey of kit of the invention in the kit
Sequence, in order to those skilled in the art's application.
Main advantages of the present invention are:
Technical solution of the present invention has good susceptibility and specially spends, and the early diagnosis of ectopic pregnancy may be implemented,
It avoids jeopardizing the generation of the serious consequences such as patient vitals and reproductive function, can greatly reduce and fail to pinpoint a disease in diagnosis risk.Simultaneously, it is also possible to make pregnant
Woman realizes self detection to ectopic pregnancy, to change the diagnostic mode and diagnostic mode of ectopic pregnancy.
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually according to conventional strip
Part such as J. Pehanorm Brooker etc. is write, Molecular Cloning:A Laboratory guide, the third edition, Science Press, condition described in 2002, or
According to the normal condition proposed by manufacturer.
The expression of embodiment 1, CNN2 in the chorionic villi of ectopic pregnancy
The present inventor obtains the uterine pregnancy chorionic villi and ectopic pregnancy chorionic villi of clinical trial object, carries out
The analysis and measurement of the otherness of gene expression.Wherein, uterine pregnancy experimental subjects 10, ectopic pregnancy experimental subjects 10.
Measurement extracts nucleic acid using conventional acquisition tissue, the method for carrying out PCR detection.
The PCR primer of use is as follows:
It is positive: 5 '-ATGAACAAGCTACAGCCGGG-3 ' (SEQ ID NO:1)；
It is reversed: 5 '-CTTAGTCTTGGCCTGCGTCA-3 ' (SEQ ID NO:2)；
The measurement result of the expression of CNN2 as shown in Figure 1, in ectopic pregnancy chorionic villi the expression of CNN2 be higher than significantly
Uterine pregnancy chorionic villi.
Embodiment 2, CNN2 plant the expression in fallopian tubal in ectopic gestation sac semina
The present inventor obtains the oviduct tissue of the embryonic implantation part of clinical ectopic pregnancy patients and far from blastaea
The part oviduct tissue of implantation carries out the analysis and measurement of CNN1, CNN2, CNN3 protein expression difference.Wherein, dystopy is pregnant
Experimental subjects of being pregnent totally 10.
Measurement is using conventional immunohistochemical staining and ELISA detection, and with anti-CNN1 antibody, (monoclonal antibody is purchased from respectively
Proteintech Group, Inc., Chicago, IL, USA), anti-CNN2 antibody (monoclonal antibody, be purchased from Novus Biologicals,
CO, USA), anti-CNN3 antibody (monoclonal antibody, be purchased from Proteintech Group, Inc., Chicago, IL, USA) detected.
The fallopian tubal part height expression of Immunohistochemical Method result such as Fig. 2, CNN2 at ectopic gestation sac semina plant.
The above results explanation, expression of the CNN2 in tubal pregnancy are related to embryonic implantation position.
The expression of CNN2 in embodiment 3, serum
The present inventor obtains ectopic pregnancy patients 84, and uterine pregnancy women 39, missed abortion 9, non-pregnant woman compares
32.The blood of these research objects is extracted, serum is obtained.CNN2 in serum is measured by the method that the port ELISA is estimated
Expression.
ELISA testing result such as table 2 prompts CNN2 that extremely significant high expression occurs in ectopic pregnancy patients serum.
Table 2
Embodiment 4, change of serum C NN2 are to the diagnostic value of ectopic pregnancy
ROC curve production: the present inventor analyzes CNN2 as single diagnosis index antidiastole ectopic pregnancy and in utero
Gestation and the diagnostic value of miscarriage.For 84 ectopic pregnancies, 39 uterine pregnancies, 9 missed abortion patients leave and take 5ml
Serum, ELISA method detection change of serum C NN2 concentration, according to CNN2 concentration and diagnostic result, SPSS statistical software are extracted after peripheral blood
Draw ROC curve.
According to the above method, ROC curve such as Fig. 3 of CNN2 Diagnosis of Ectopic Pregnancy is obtained.Sensitivity reaches 95.2%, specifically
Degree reaches 93.7%.
The above results explanation has good sensitivity and specificity using change of serum C NN2 as diagnosis index, and CNN2 is
A kind of diagnosis marker of good clinical ectopic pregnancy.
Embodiment 5, change of serum C NN2 are in the postoperative expression of ectopic pregnancy
The present inventor obtains clinically 84 ectopic pregnancy patients, respectively in the preoperative and postoperative their serum of detection
CNN2 is horizontal.
The measurement result of preoperative and postoperative change of serum C NN2 level is as shown in figure 4, preoperative 91.55 ± 21.89ng/ml, art
78.99 ± 25.34ng/ml afterwards.As it can be seen that change of serum C NN2 is lowered in the postoperative expression that conspicuousness occurs of ectopic pregnancy.
The correlation of embodiment 6, change of serum C NN2 concentration and pregnant week
The present inventor obtains clinically 84 ectopic pregnancy patients, extracts serum, ELISA method inspection after leaving and taking 5ml peripheral blood
Change of serum C NN2 concentration is surveyed, and calculates patient's pregnant week, SPSS statistical software analyzes CNN2 concentration and pregnant week correlation.
As a result such as Fig. 5, abscissa indicate that pregnancy number of days, ordinate indicate CNN2 concentration, and each point represents a specific disease
Example.It can be seen that CNN2 concentration and pregnant week non-correlation.
To sum up, the expression to CNN2 in ectopic pregnancy, the sensitivity of change of serum C NN2 Diagnosis of Ectopic Pregnancy and specificity,
Variation and CNN2 concentration of the CNN2 before and after Operation of Ectopic Pregnancy are analyzed with menopause day data/coherency.As can be seen that
The sensitivity of CNN2 Diagnosis of Ectopic Pregnancy and specificity are apparently higher than currently used β-hCG and progesterone, and accuracy significantly improves；
Meanwhile compared with β-hCG and progesterone, it is more suitable that CNN2 and pregnant week and internal sex hormone level are unrelated therefore relative constant
Ectopic pregnancy specific serum marker.
All references mentioned in the present invention is incorporated herein by reference, independent just as each document
It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can
To make various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims
It encloses.
Sequence table
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Claims (10)
- The purposes of 1.Calponin-h2 albumen or its encoding gene in the diagnostic reagent or kit of preparation ectopic pregnancy.
- 2. purposes as described in claim 1, which is characterized in that the ectopic pregnancy are as follows: tubal pregnancy.
- 3. purposes as described in claim 1, which is characterized in that the diagnostic reagent is selected from:The primer of the encoding gene of specific amplification Calponin-h2 albumen；OrThe encoding gene of specific recognition Calponin-h2 albumen or the probe of its transcript；OrSpecifically bind the binding molecule of Calponin-h2 albumen；OrThe encoding gene of specific recognition Calponin-h2 albumen or the chip of its transcript.
- 4. purposes as claimed in claim 3, which is characterized in that the coding of the specific amplification Calponin-h2 albumen The primer of gene is primer pair, and nucleotide sequence is as shown in SEQ ID NO:1 and SEQ ID NO:2.
- 5. purposes as claimed in claim 3, which is characterized in that the combination of the specific binding Calponin-h2 albumen Molecule is antibody.
- 6. purposes as claimed in claim 5, which is characterized in that the antibody is monoclonal antibody or polyclonal antibody.
- 7. purposes as described in claim 1, which is characterized in that the diagnostic reagent is the diagnosis using serum as sample to be tested Reagent, using blood plasma as the diagnostic reagent of sample to be tested or using peripheral blood mononuclear cells as the diagnostic reagent of sample to be tested；Preferably Ground is the diagnostic reagent using serum as sample to be tested.
- 8. a kind of kit for Diagnosis of Ectopic Pregnancy, which is characterized in that contain in the kit: detection The diagnostic reagent of the expression or expression quantity of Calponin-h2 albumen or its encoding gene, or be mounted with and change diagnostic reagent Solid phase carrier；Preferably, the diagnostic reagent is selected from: the primer of the encoding gene of specific amplification Calponin-h2 albumen；Or it is special The encoding gene of opposite sex identification Calponin-h2 albumen or the probe of its transcript；Or specific binding Calponin-h2 albumen Binding molecule；Specific recognition Calponin-h2 albumen encoding gene or its transcript chip；Preferably, the diagnostic reagent is the primer of the encoding gene of specific amplification Calponin-h2 albumen, nucleotide Sequence is as shown in SEQ ID NO:1 and SEQ ID NO:2；Preferably, in the kit further include: nucleic acid extraction reagent；Pcr reagent；The examination of protein immunization groupization Agent；Enzyme chain immune response reagent；Or operation instructions.
- 9. a kind of method for the diagnostic reagent for preparing ectopic pregnancy, which is characterized in that the described method includes: with Calponin-h2 Albumen or its encoding gene are object, prepare specific recognition or in conjunction with the Calponin-h2 albumen or its encoding gene Reagent；Preferably, the reagent includes:The primer of the encoding gene of specific amplification Calponin-h2 albumen；OrThe encoding gene of specific recognition Calponin-h2 albumen or the probe of its transcript；OrSpecifically bind the binding molecule of Calponin-h2 albumen；OrThe encoding gene of specific recognition Calponin-h2 albumen or the chip of its transcript.
- 10. method as claimed in claim 9, which is characterized in that the method also includes: by the specific recognition or knot On the load reagents to solid phase carrier for closing the Calponin-h2 albumen or its encoding gene；Preferably, the solid phase carrier It include: test paper or test strips, microballoon, slide.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112034180A (en) * | 2020-08-18 | 2020-12-04 | 四川大学华西第二医院 | Use and product of keratin 1 in ectopic pregnancy |
Citations (1)
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| CN102272325A (en) * | 2008-11-17 | 2011-12-07 | 威拉赛特公司 | Methods and compositions of molecular profiling for disease diagnostics |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102272325A (en) * | 2008-11-17 | 2011-12-07 | 威拉赛特公司 | Methods and compositions of molecular profiling for disease diagnostics |
Non-Patent Citations (2)
| Title |
|---|
| RUIQINGZHANGM.D等: "Serum calponin 2 is a novel biomarker of tubal ectopic pregnancy", 《FERTILITY AND STERILITY》 * |
| 谷永红等: "Calponin-1表达沉默对子宫平滑肌细胞生物学行为的影响", 《南方医科大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112034180A (en) * | 2020-08-18 | 2020-12-04 | 四川大学华西第二医院 | Use and product of keratin 1 in ectopic pregnancy |
| CN112034180B (en) * | 2020-08-18 | 2021-09-24 | 四川大学华西第二医院 | Use and product of keratin 1 in ectopic pregnancy |
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| CN109971847B (en) | 2022-08-02 |
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