CN1495265A - Definition of Gardner's bacillus DNA sequence (1) specificity of vagina and its application - Google Patents
Definition of Gardner's bacillus DNA sequence (1) specificity of vagina and its application Download PDFInfo
- Publication number
- CN1495265A CN1495265A CNA021334064A CN02133406A CN1495265A CN 1495265 A CN1495265 A CN 1495265A CN A021334064 A CNA021334064 A CN A021334064A CN 02133406 A CN02133406 A CN 02133406A CN 1495265 A CN1495265 A CN 1495265A
- Authority
- CN
- China
- Prior art keywords
- sequence
- bacillus
- denashi
- gardner
- specificity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to the definition of specificity of a tract of vaginal Gardner's bacillus DNA sequence (I) and its application. It utilizes the search and analysis of gene data base material to define the specificity of at tract of vaginal Gardern's bacillus DNA sequence (I) and PCR amplification primer, and the length of said sequence is 200bp. It utilizes the PCR amplification and clone of woman vaginal Gardner's bacillus infection specimen to obtain said sequence. The sequencing shows that said sequence is identical to the predefined sequence, and the results of biological informatic method analysis and gene chip hybridization analysis show the specificity of said sequence. Said invention can be used as probe of gene chip diagnosis and PCR amplification detection sequence for diagnosing vaginal Gardner's bacillus.
Description
Technical field:
The present invention relates to the specific definite and application of one section vagina Jia Denashi bacillus (Gardnerella vaginalis) dna sequence dna (1), belong to field of biomedicine technology.
Background technology:
Vagina Jia Denashi bacillus (Gardnerella vaginalis) is the pathogen that a kind of woman vagina often infects, and diagnosis accurately is to carry out the foundation of treatment immediately and reliably, can only diagnose according to clinical indication at present.PCR and gene chip diagnosis are a kind of rapid and precise detection methods.And the diagnostic method of this class depends on the special nucleotide sequence of acquisition.
Summary of the invention:
The invention reside in by information biology and Protocols in Molecular Biology, determined one section vagina Jia Denashi bacillus (Gardnerella vaginalis) specific nucleotide sequence and PCR primer thereof.This sequence can be used for setting up gene chip with corresponding PCR primer and PCR method is carried out the diagnosis of vagina Jia Denashi bacillus (Gardnerella vaginalis).
Technical scheme of the present invention is: the present invention utilizes existing method to determine one section vagina Jia Denashi bacillus (Gardnerella vaginalis) specific DNA sequence (1) and the PCR primer thereof of length for 200bp, carry out pcr amplification with primer, its sequence is as follows:
ggcatgagaa?aagtggctgt?tccatttgga?gtaagccaaa?ttgcacaagt?ggctgccatt 60
gcctctttgg?aacctcaagt?gcttgtagag?atgcgcaacc?gcgtagatgc?acttgttaag 120
gagcgtaatc?gcgttgtaag?cggtttgcgt?gaacaagggt?ggaatatcgt?cgagccgtac 180
gcaaacttct?tctggatgcc 200
Primer sequence is:
1.5’-ggcatgagaaaagtggctgt-3’
2.5’-ggcatccagaagaagtttgc-3’
Above-mentioned vagina Jia Denashi bacillus (Gardnerella vaginalis) specific DNA sequence (1) is used to diagnose the probe sequence of the gene chip of vagina Jia Denashi bacillus, and carry out pcr amplification by above-mentioned primer sequence, hybridize behind the mark and detect or be directly used in the PCR diagnosis.
The employed method of setting up with the present invention of clinical detection technique has quick, accurate, easy and simple to handle, low cost and other advantages.
Description of drawings:
Fig. 1 is the pcr amplification collection of illustrative plates of clinical samples.
Fig. 2, Fig. 2 are continuous to be sequencer map.Wherein Fig. 2 be sequence, Fig. 2 of measuring continuous be the primitive sequencer collection of illustrative plates.
Fig. 3 is with the sequencing result and the comparative analysis of estimating sequence of Blast software to pcr amplification product.
Fig. 4, Fig. 4 are continuous to be Blast Search Results in the gene database.
Fig. 5 is the gene chip hybridization result.
Embodiment:
Be keyword at first with vagina Jia Denashi bacillus (Gardnerella vaginalis), the nucleotide sequence of search vagina Jia Denashi bacillus (Gardnerella vaginalis) in gene database.Obtain the closer nucleotide sequence of several relations altogether, with these nucleotide sequences with the nucleotide sequence (nr) in the online use software of the online Blast of the NCBI comparison gene database, discharge nonspecific nucleotide sequence, obtained the specific nucleotide sequence of 1Kb.With this section sequences Design surplus 20 to the PCR primer, extract DNA with clinical samples, carry out pcr amplification, screen a pair of sequence and the primer that is fit to carry out pcr amplification, the product that pcr amplification obtains is consistent with the expectation size, and (see Fig. 1, sequence number 1 is standard molecular weight DNA among the figure, the molecular weight size is respectively from top to bottom: 622,527,404,309,242/238,217,201,190,180,160+160,147+147,123; 2.4.5 be clinical negative sample, there is not amplified production; 3. clinical positive sample, amplified production is very pure, and size about 200 is consistent with the expectation size.The collection of illustrative plates result shows with the special amplification clinical sample of the energy of the primer among the present invention), clone with T-vector then, and the order-checking (sequencer address is seen Fig. 2, Fig. 2 is continuous, among the figure since 59, to 258,200bp is a cloned sequence altogether, all the other sequences are the cloning vector sequence), sequence is 200bp, estimate sequence by the comparison of Blast software, demonstration is consistent with the sequence of estimating (to be seen Fig. 3, removes the 24th among the figure, 45 and 111 change over g by a, and the 80th and 170 changes over c by t, the 99th, 168 and 180 are changed over outside the t by c, all the other are all consistent with the expectation sequence), show that this sequence can be can be used for vagina Jia Denashi bacillus (Gardnerella vagihalis) PCR and detect by specific amplification.Sequence is with DNA (the comprising cDNA) sequence of the Blast muca gene database of NCBI website, this sequence is only consistent with the sequence of vagina Jia Denashi bacillus (Gardnerella vaginalis), and do not have homology [to see that Fig. 4, Fig. 4 are continuous with other species sequence, search result shows among the figure, plasmid dna sequence and this sequence homology of having only vagina Jia Denashi bacillus (Gardnerella vaginalis), and be to match fully, the sequence homology that one section about 70bp is arranged also is the sequence of vagina Jia Denashi bacillus (Gardnerellavaginalis).And other sequence has only the short sequence about 40 characters, because these sequences do not have primer to match therewith, so can not be detected in detection].With behind this sequence mark as probe with contain the gene chip hybridization of 45 gene locuss of 9 kinds of kinds of genitourinary infection pathogen, have only sequence of the present invention that hybridization signal is arranged therewith, and other all sequences therewith the amixia signal (see Fig. 5, be 4 repetitions among the figure, each repeats to contain 9 kinds of urogenital infections disease pathogen things totally 45 different sites and 4 control site.Results of hybridization shows to have only this sequence that special signal is arranged, and other equal amixia signal), show that this sequence is the distinguished sequence of vagina Jia Denashi bacillus (Gardnerella vaginalis).
The present invention can be used for the clinical detection that PCR method or method for gene chip carry out vagina Jia Denashi bacillus (Gardnerellavaginalis).Carry out pcr amplification with sequence provided by the invention, obtain product according to PCR method and whether occur judging whether to infect into vagina Jia Denashi bacillus (Gardnerella vaginalis) with size.An also available sequence is a probe, as a detection site in the gene chip, then with the amplification of the PCR primer sequence among the present invention clinical samples, and hybridizes detection behind the mark.
The specification sheets sequence table
＜110〉Kunming Huanji Biochip Development Co., Ltd
＜120〉one section vagina Jia Denashi bacillus DNA sequence (1) is specific determines
<140>
<141>
<160>1
<210>1
<211>200
<212>DNA
＜213〉vagina Jia Denashi bacillus (Gardnerella vaginalis)
<220>
<221>gene
<222>(1)…(200)
<400>1
ggcatgagaa?aagtggctgt?tccatttgga?gtaagccaaa?ttgcacaagt?ggctgccatt 60
gcctctttgg?aacctcaagt?gcttgtagag?atgcgcaacc?gcgtagatgc?acttgttaag 120
gagcgtaatc?gcgttgtaag?cggtttgcgt?gaacaagggt?ggaatatcgt?cgagccgtac 180
gcaaacttct?tctggatgcc 200
Claims (2)
1. one section vagina Jia Denashi bacillus DNA sequence (1) is specific determines that the length that it is characterized in that this sequence is 200bp, and its sequence and PCR primer are as follows:
ggcatgagaa?aagtggctgt?tccatttgga?gtaagccaaa?ttgcacaagt?ggctgccatt 60
gcctctttgg?aacctcaagt?gcttgtagag?atgcgcaacc?gcgtagatgc?acttgttaag 120
gagcgtaatc?gcgttgtaag?cggtttgcgt?gaacaagggt?ggaatatcgt?cgagccgtac 180
gcaaacttct?tctggatgcc 200
Primer sequence is:
1.5’-ggcatgagaaaagtggctgt-3’
2.5’-ggcatccagaagaagtttgc-3’
2. the specific application of one section vagina Jia Denashi bacillus DNA sequence (1), it is characterized in that this sequence is used to diagnose the probe sequence of the gene chip of vagina Jia Denashi bacillus, and carry out pcr amplification by above-mentioned primer sequence, hybridize behind the mark and detect or be directly used in the PCR diagnosis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA021334064A CN1495265A (en) | 2002-07-02 | 2002-07-02 | Definition of Gardner's bacillus DNA sequence (1) specificity of vagina and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA021334064A CN1495265A (en) | 2002-07-02 | 2002-07-02 | Definition of Gardner's bacillus DNA sequence (1) specificity of vagina and its application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1495265A true CN1495265A (en) | 2004-05-12 |
Family
ID=34231306
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA021334064A Pending CN1495265A (en) | 2002-07-02 | 2002-07-02 | Definition of Gardner's bacillus DNA sequence (1) specificity of vagina and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1495265A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104975094A (en) * | 2015-07-13 | 2015-10-14 | 王爱玲 | Kit of gardnerella vaginalis for diagnosing colpitis |
| CN105087812B (en) * | 2015-07-13 | 2016-10-12 | 北京博奥医学检验所有限公司 | A kind of bacterial vaginitis detection kit |
-
2002
- 2002-07-02 CN CNA021334064A patent/CN1495265A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104975094A (en) * | 2015-07-13 | 2015-10-14 | 王爱玲 | Kit of gardnerella vaginalis for diagnosing colpitis |
| CN105154570B (en) * | 2015-07-13 | 2016-06-08 | 武汉迪安医学检验所有限公司 | A kind of test kit for diagnosing colpitic gardnerella vaginalis |
| CN105087812B (en) * | 2015-07-13 | 2016-10-12 | 北京博奥医学检验所有限公司 | A kind of bacterial vaginitis detection kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103045716B (en) | Method for detecting isoniazid drug resistance of mycobacterium tuberculosis by using pyrosequencing technology | |
| CN1495265A (en) | Definition of Gardner's bacillus DNA sequence (1) specificity of vagina and its application | |
| CN1206366C (en) | Determination of man-shaped mycoplasma DNA sequence (2) specificity and its use | |
| CN1285734C (en) | Determination of one section human type mycoplasmas DNA sequence (1) specificity and its application | |
| CN1238521C (en) | Determination of a section of man-shaped mycoplasma DNA sequence (4) specificity and its use | |
| CN1168835C (en) | Determination for specificity of Candida albicans DNA sequence (2) and use thereof | |
| CN1205341C (en) | Determination of man-shaped mycoplasma DNA sequence (5) specificity and its use | |
| CN1206367C (en) | Determination of a section of mycoplasma urealyticum DNA sequence (4) specificity and its use | |
| CN1493698A (en) | Determination of one section vagina bacillus Gardner DNA sequence (3) specificity and its application | |
| CN1170945C (en) | Determination for specificity of ureaplasma DNA sequence (1) and use thereof | |
| CN1168833C (en) | Determination for specificity of candida albicans DNA sequence (4) and use thereof | |
| CN1168834C (en) | Determination for specificity of Candida albicans DNA sequence (5) and use thereof | |
| CN1168836C (en) | Determination for specificity of candida albicans DNA sequence (3) and use thereof | |
| CN1570136A (en) | Human type mycoplasma DNA sequence (3) specificity determination and its uses | |
| CN1267564C (en) | Determination of one section urea degraded mycoplasmos DNA sequence (5) specificity and its application | |
| CN1232650C (en) | Specificity determination and application of one section of gonococcus DNA sequence | |
| CN1495267A (en) | Definition of human papillomavirus-6 DNA sequence (4) specificity and its application | |
| CN1495269A (en) | Definition of vaginal Gardner's bacillus DNA sequence (4) specificity and its application | |
| CN1495264A (en) | Definition of Gardner's bacillus DNA sequence (5) specificity of vagina and its application | |
| CN1493699A (en) | Determination of one section vagina bucillus Gardner DNA sequence (2) specificity and its application | |
| CN113718053A (en) | Probe and primer pair for detecting yersinia sporogenes, detection method and application | |
| CN1495266A (en) | Definition of human papillomavirus-6 DNA squence (5) speicificity and its application | |
| CN1495268A (en) | Definition of human papillomavirus-6 DNA sequence (3) specificity and its application | |
| CN1495270A (en) | Definition of human papillomavirus-6 DNA sequence (2) specificity and its application | |
| CN1495271A (en) | Definition of human papillomavirus-6 DNA sequence (1) specificity and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C57 | Notification of unclear or unknown address | ||
| DD01 | Delivery of document by public notice |
Addressee: Zhu Wenzhi Document name: Notice of first review |
|
| C57 | Notification of unclear or unknown address | ||
| DD01 | Delivery of document by public notice |
Addressee: Zhu Wenzhi Document name: Deemed as a notice of withdrawal (Trial) |
|
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |