CN102363816A - Reagent for detecting enterohemorrhagic escherichia coli by using cross primer nucleic acid isothermal amplification technology, amplification method and detection method for enterohemorrhagic escherichia coli - Google Patents
Reagent for detecting enterohemorrhagic escherichia coli by using cross primer nucleic acid isothermal amplification technology, amplification method and detection method for enterohemorrhagic escherichia coli Download PDFInfo
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- CN102363816A CN102363816A CN2011103827633A CN201110382763A CN102363816A CN 102363816 A CN102363816 A CN 102363816A CN 2011103827633 A CN2011103827633 A CN 2011103827633A CN 201110382763 A CN201110382763 A CN 201110382763A CN 102363816 A CN102363816 A CN 102363816A
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Abstract
The invention discloses a reagent for detecting enterohemorrhagic escherichia coli by using a cross primer nucleic acid isothermal amplification technology, an amplification method and a detection method for enterohemorrhagic escherichia coli. Five primer group sequences are designed, namely 5'CAGTCGTACGGGGATGCAG, 5'TCAGCAGTCATTACATAAG, 5'GCTCTTGCCACAGACTGCGTC, 5'AGGTTCCGCTATGCGACAT and 5'GCTCTTGCCACAGACTGCGTCAGTTTCGCCATTCGTTGACTACTT. Aiming at the enterohemorrhagic escherichia coli O157:H7 bacteria, the defects in the prior art are overcome by the reagent, and a method for detecting the enterohemorrhagic escherichia coli quickly at low cost by using the cross primer nucleic acid isothermal amplification technology is provided.
Description
Technical field
The present invention relates to the bacteriologic test technology, specifically use cross primer nucleic acid constant-temperature amplification technology for detection EHEC O157:H7 reagent and amplification method and detection method.
Background technology
EHEC O157:H7 belongs to the enterobacteriaceae Escherichia, is the main serotype of EHEC, the hemorrhagic colitis that can cause.Nineteen eighty-two is reined in Gang Zhou and Michigan State in U.S. Russia, and is separated and name in patient's ight soil of the food poisoning that causes because of edible hamburger.According to the Center for Disease Control (CDC) report, 20,000 many cases patients are found in the annual whole nation approximately.Death toll 200-500 people, five big states more than 20 country comes to light or causes and break out with popular in the whole world successively later on.Found so far from 82 years; It is the last ten-days period in May, 97 that the biggest one outburst takes place, and 6 mass food poisoning incidents take place in succession for tens middle schools, county such as Japanese Okayama, Hiroshima and kindergarten, and number is 1600 people nearly; Cause 3 death of child, hospitalized child reaches 80 people.Japanese celestial platform and deer island form the poisoning number and cross ten thousand people simultaneously, dead 11 people, and the burst that involves 44 Dou Fu counties is poisoned by food, and causes global concern.Countries and regions such as the U.S., Canada, Sweden, Australia, sco, Wales have also been reported sporadic infection and outbreak of epidemic in succession in addition.China was separated to EHEC O157:H7 in 1988 first.See that from existing epidemiology survey data also there is Sporadic cases in China, also do not have the report of eruption and prevalence.Also through monitoring, successively, found EHEC O157:H7 in the food such as meat in recent years from ox, pig, sheep, ight soil.Accurate detection EHEC O157:H7 is the precondition that effectively prevention and control enterobacteria infect.Along with development of molecular biology, the method for determining bacteria in the real work on traditional simple biochemical test level to molecular biological method such as technical developments such as PCR, probe hybridization.The cross primer nucleic acid constant-temperature amplification technology that recent development is come out is a kind of molecular detecting method more efficiently, by the approval of a lot of countries, and greatly develops.
Summary of the invention
To EHEC O157:H7; The present invention has overcome shortcoming of the prior art, provide a kind of quick, convenient, cheaply use cross primer nucleic acid constant-temperature amplification technology for detection EHEC O157:H7 reagent and amplification method and detection method.
In order to solve the problems of the technologies described above, the technical scheme that the present invention adopts is: a kind of reagent that uses cross primer nucleic acid constant-temperature amplification technology for detection EHEC comprises following five primers:
SEQ?ID?NO.1：5’?CAGTCGTACGGGGATGCAG
SEQ?ID?NO.2：5’?TCAGCAGTCATTACATAAG
SEQ?ID?NO.3：5’?GCTCTTGCCACAGACTGCGTC
SEQ?ID?NO.4：5'?AGGTTCCGCTATGCGACAT
SEQ?ID?NO.5：5'?GCTCTTGCCACAGACTGCGTCAGTTTCGCCATTCGTTGACTACTT
Wherein 5 ' of primer sequence 3 end has fluorescein isothiocyanate group mark, and 5 ' end of primer sequence 4 has vitamin H fluorophor mark.
Each component composition is following in the nucleic acid amplification reaction system:
Constituent concentration application of sample amount
Bst enzyme 5U/ μ L 1 μ L
Bst enzyme buffer liquid-5 μ L
SEQ?ID?NO.1： 10μmol/L 1μL
SEQ?ID?NO.2： 10μmol/L 1μL
SEQ?ID?NO.3： 10μmol/L 0.5μL
SEQ?ID?NO.4： 10μmol/L 0.5μL
SEQ?ID?NO.5： 10μmol/L 0.2μL
Distilled water 13.8 μ L
TV 25 μ L
Wherein the composition of Bst enzyme buffer liquid is 20 mM Tris-HCl, 10 mM (NH
4)
2SO
4, 10 mM KCl, 2 mM MgSO
4, mass percent 0.1 % Triton X-100, and pH is 8.8.
The amplification method of the reagent of described utilization cross primer nucleic acid constant-temperature amplification technology for detection EHEC, the nucleic acid amplification program is:
(1) 63 ℃ 90 minutes;
(2) 80 ℃ 2 minutes.
The detection method of the reagent of described utilization cross primer nucleic acid constant-temperature amplification technology for detection EHEC; The reaction drop extremely can be detected on fluorescein isothiocyanate group and the colloidal gold test of vitamin H fluorophor with the nucleic acid samples of tense marker; In 15-30 minute, read the result; If only a red line appears in C in the Quality Control district, no EHEC O157:H7 in the expression sample; If two red lines, a detection line, there is EHEC O157:H7 in a nature controlling line in the expression sample; If not having red line occurs.Show and detect failure, sample needs to detect again.
Compared with prior art; The invention has the beneficial effects as follows: compare traditional Physiology and biochemistry detection method; Cross primer nucleic acid constant-temperature amplification technology is applicable to directly from patient's mycetome liquid, ight soil; Whether amplified target gene in the clinical samples such as food and body fluid culture only needs a constant-temperature amplification just can detect EHEC O157:H7 and exists, and has improved efficient greatly and has practiced thrift the time.Other method of comparing, our legal system only need simple equipment to get final product, and have improved cost performance greatly and have practiced thrift cost.This technology for detection EHEC O157:H7 have detect accurately, high specificity, highly sensitive characteristics, can identify quickly and accurately, avoided cultivating repeatedly, save time; This method does not receive the influence of culture condition and bacterium physiological status, and is more accurate than the Physiology and biochemistry authentication method.
Description of drawings
Fig. 1 is the synoptic diagram of interpretation as a result of the present invention.
Fig. 2 is experimental result (the detector bar 1. enteritis EHEC O157:H7 CIQ103 detected results that the present invention uses cross primer nucleic acid constant-temperature amplification technology for detection EHEC O157:H7; Detector bar 2. enteritis EHEC O157:H7 CIQ109 detected results; Detector bar 3. enteritis EHEC O157:H7 CIQ79 detected results; Detector bar 4. Shigellae ATCC12022 detected results; Detector bar 5. blanks).
Embodiment
Below in conjunction with accompanying drawing and embodiment the present invention is done further explain:
The present invention realizes through following technical scheme:
(1) the designs specificity Oligonucleolide primers is used for cross primer nucleic acid constant-temperature amplification technology for detection;
(2) integrate each primer and make it non-interference.
(3) with the primer sequence group, amplification testing sample template is carried out the specific amplification of goal gene;
(4) can be after amplification finishes through specific colloidal gold colloidal gold detection test paper strip visual observation.
This method comprises: use this method and detect employed primer sets of EHEC O157:H7 and supporting with it nucleic acid amplification reaction condition.Wherein primer sequence is following:
SEQ?ID?NO.1：5’?CAGTCGTACGGGGATGCAG
SEQ?ID?NO.2：5’?TCAGCAGTCATTACATAAG
SEQ?ID?NO.3：5’?GCTCTTGCCACAGACTGCGTC
SEQ?ID?NO.4：5'?AGGTTCCGCTATGCGACAT
SEQ?ID?NO.5：5'?GCTCTTGCCACAGACTGCGTCAGTTTCGCCATTCGTTGACTACTT
Wherein 5 ' of primer sequence 3 end has fluorescein isothiocyanate group mark.
5 ' end of primer sequence 4 has vitamin H fluorophor mark.
This primer is selected design for congratulating (shiga-like toxin 1 subunit A) in the appearance enterotoxin A subunit gene with the pathogenic closely-related will of EHEC, and reference sequences is GenBank AE005174.2
Escherichia coliPartial stx1 gene for shiga toxin 1, strain EHEC FE94076, detecting gene region is 2996033 ~ 2996980.
Among preferred the present invention in the fluorescent PCR reaction system each component composition following:
Constituent concentration application of sample amount
Bst enzyme 5U/ μ L 1 μ L
Bst enzyme buffer liquid-5 μ L
SEQ?ID?NO.1： 10μmol/L 1μL
SEQ?ID?NO.2： 10μmol/L 1μL
SEQ?ID?NO.3： 10μmol/L 0.5μL
SEQ?ID?NO.4： 10μmol/L 0.5μL
SEQ?ID?NO.5： 10μmol/L 0.2μL
Distilled water 13.8 μ L
TV 25 μ L
Wherein the composition of Bst enzyme buffer liquid is 20 mM Tris-HCl, 10 mM (NH
4)
2SO
4, 10 mM KCl, 2 mM MgSO
4, mass percent 0.1 % Triton X-100, and pH is 8.8.
The nucleic acid amplification program is:
(1) 63 ℃ 90 minutes;
(2) 80 ℃ 2 minutes.
Like Fig. 1, shown in 2; Nucleic acid amplification product can detect through the colloidal gold test that can detect the nucleic acid amplification mark, and decision method is following: will react drop to detecting on fluorescein isothiocyanate group and the colloidal gold test of vitamin H fluorophor with the nucleic acid samples of tense marker, and in 15-30 minute, read the result; As only a red line appears in C in the Quality Control district; No EHEC O157:H7 in the expression sample, if two red lines, a detection line; Article one, there is EHEC O157:H7 in nature controlling line in the expression sample.Do not occur if there is red line, show and detect failure, sample needs to detect again.
Sample: certain fecal sample.
Detect the doubtful bacterium colony of EHEC O157:H7 with conventional physiology, biochemical method, carry out following cross primer nucleic acid constant-temperature amplification then and detect:
(1) getting 0.1 gram sample to be checked suspends with 1 gram saline water.
(2) get 1 gram sample and carry out the nucleic acid extracting.
(3) cross primer nucleic acid constant-temperature amplification
Each component composition is following in the reaction system:
Constituent concentration application of sample amount
Bst enzyme 5U/ μ L 1 μ L
Bst enzyme buffer liquid-5 μ L
SEQ?ID?NO.1： 10μmol/L 1μL
SEQ?ID?NO.2： 10μmol/L 1μL
SEQ?ID?NO.3： 10μmol/L 0.5μL
SEQ?ID?NO.4： 10μmol/L 0.5μL
SEQ?ID?NO.5： 10μmol/L 0.2μL
Distilled water 13.8 μ L
TV 25 μ L
Wherein the composition of Bst enzyme buffer liquid is 20 mM Tris-HCl, 10 mM (NH
4)
2SO
4, 10 mM KCl, 2 mM MgSO
4, mass percent 0.1 % Triton X-100, and pH is 8.8.
The nucleic acid amplification program is:
63 ℃ 90 minutes; 80 ℃ 2 minutes.
Carry out 3 pipe experiments altogether, what wherein the DNA sample was added is respectively: EHEC O157:H7DNA template standard article; This testing sample DNA; The negative control of no DNA.
(4) result observes
The reaction drop extremely can be detected on fluorescein isothiocyanate group and the colloidal gold test of vitamin H fluorophor with the nucleic acid samples of tense marker; In 20 minutes, read the result; EHEC O157:H7DNA template standard article two red lines occur with this testing sample DNA;, a detection line, a nature controlling line; Only a red line appears to the negative control of no DNA in C in the Quality Control district.There is EHEC O157:H7 in comprehensive above result in the expression testing sample.
After 3 days, prove that the purpose bacterium colony of being checked is EHEC O157:H7 through conventional microorganism culturing and biochemistry detection.Cross primer nucleic acid constant-temperature amplification assay is consistent with the biochemistry detection result.
Sample: certain ground pork.
Detect the doubtful bacterium colony of EHEC O157:H7 with conventional physiology, biochemical method, carry out following cross primer nucleic acid constant-temperature amplification then and detect:
(1) gets 100 grams sample to be checked, pulverize.
(2) get 1 gram sample and carry out the nucleic acid extracting.
(3) cross primer nucleic acid constant-temperature amplification
Constituent concentration application of sample amount
Bst enzyme 5U/ μ L 1 μ L
Bst enzyme buffer liquid-5 μ L
SEQ?ID?NO.1： 10μmol/L 1μL
SEQ?ID?NO.2： 10μmol/L 1μL
SEQ?ID?NO.3： 10μmol/L 0.5μL
SEQ?ID?NO.4： 10μmol/L 0.5μL
SEQ?ID?NO.5： 10μmol/L 0.2μL
Distilled water 13.8 μ L
TV 25 μ L
Wherein the composition of Bst enzyme buffer liquid is 20 mM Tris-HCl, 10 mM (NH
4)
2SO
4, 10 mM KCl, 2 mM MgSO
4, mass percent 0.1 % Triton X-100, and pH is 8.8.
The nucleic acid amplification program is:
63 ℃ 90 minutes; 80 ℃ 2 minutes.
Carry out 3 pipe experiments altogether, what wherein the DNA sample was added is respectively: EHEC O157:H7DNA template standard article; This testing sample DNA; The negative control of no DNA.
(4) result observes
The reaction drop extremely can be detected on fluorescein isothiocyanate group and the colloidal gold test of vitamin H fluorophor with the nucleic acid samples of tense marker; In 20 minutes, read the result; EHEC O157:H7DNA template standard article two red lines occur with this testing sample DNA;, a detection line, a nature controlling line; Only a red line appears to the negative control of no DNA in C in the Quality Control district.There is EHEC O157:H7 in comprehensive above result in the expression testing sample.
After 3 days, prove that the purpose bacterium colony of being checked is EHEC O157:H7 through conventional microorganism culturing and biochemistry detection.Cross primer nucleic acid constant-temperature amplification assay is consistent with the biochemistry detection result.
Above-described embodiment only is used to explain technological thought of the present invention and characteristics; Its purpose is to make those skilled in the art can understand content of the present invention and implements according to this; Can not only limit claim of the present invention with present embodiment; Be equal variation or the modification that all disclosed spirit is done, still drop in the claim of the present invention.
Claims (4)
1. a reagent that uses cross primer nucleic acid constant-temperature amplification technology for detection EHEC is characterized in that, comprises following five primers:
SEQ?ID?NO.1：5’?CAGTCGTACGGGGATGCAG
SEQ?ID?NO.2：5’?TCAGCAGTCATTACATAAG
SEQ?ID?NO.3：5’?GCTCTTGCCACAGACTGCGTC
SEQ?ID?NO.4：5'?AGGTTCCGCTATGCGACAT
SEQ?ID?NO.5：5'?GCTCTTGCCACAGACTGCGTCAGTTTCGCCATTCGTTGACTACTT
Wherein 5 ' of primer sequence 3 end has fluorescein isothiocyanate group mark, and 5 ' end of primer sequence 4 has vitamin H fluorophor mark.
2. the reagent of utilization cross primer nucleic acid constant-temperature amplification technology for detection EHEC according to claim 1 is characterized in that each component composition is following in the nucleic acid amplification reaction system:
Constituent concentration application of sample amount
Bst enzyme 5U/ μ L 1 μ L
Bst enzyme buffer liquid-5 μ L
SEQ?ID?NO.1： 10μmol/L 1μL
SEQ?ID?NO.2： 10μmol/L 1μL
SEQ?ID?NO.3： 10μmol/L 0.5μL
SEQ?ID?NO.4： 10μmol/L 0.5μL
SEQ?ID?NO.5： 10μmol/L 0.2μL
DNA sample 2 μ L
Distilled water 13.8 μ L
TV 25 μ L
Wherein the composition of Bst enzyme buffer liquid is 20 mM Tris-HCl, 10 mM (NH
4)
2SO
4, 10 mM KCl, 2 mM MgSO
4, mass percent 0.1 % Triton X-100, and pH is 8.8.
3. the amplification method of the reagent of the described utilization cross primer of claim 1 a nucleic acid constant-temperature amplification technology for detection EHEC is characterized in that the nucleic acid amplification program is:
(1) 63 ℃ 90 minutes;
(2) 80 ℃ 2 minutes.
4. the detection method of the reagent of the described utilization cross primer of claim 1 a nucleic acid constant-temperature amplification technology for detection EHEC; It is characterized in that; The reaction drop extremely can be detected on fluorescein isothiocyanate group and the colloidal gold test of vitamin H fluorophor with the nucleic acid samples of tense marker; In 15-30 minute, read the result, if only a red line appears in C in the Quality Control district, no EHEC O157:H7 in the expression sample; If two red lines, a detection line, there is EHEC O157:H7 in a nature controlling line in the expression sample; Do not occur if there is red line, show and detect failure, sample needs to detect again.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103343164A (en) * | 2013-06-17 | 2013-10-09 | 武汉轻工大学 | Multiplex PCR detection method and kit for Shiga toxin-producing Escherichia coli and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101343661A (en) * | 2008-08-22 | 2009-01-14 | 华南理工大学 | Constant temperature nucleic acid amplification fast detecting reagent kit for bacillus coli O157 and use method thereof |
| CN101487057A (en) * | 2009-01-23 | 2009-07-22 | 浙江省疾病预防控制中心 | Loop-mediated isothermal amplification fast detecting reagent kit and method for O157:H7 coliform |
| CN101864494A (en) * | 2010-04-13 | 2010-10-20 | 上海国际旅行卫生保健中心 | Constant-temperature amplification detection kit of A(H1N1) virus and detection method thereof |
| CN101899501A (en) * | 2010-02-10 | 2010-12-01 | 江苏出入境检验检疫局动植物与食品检测中心 | Constant temperature amplification detection kit and method for detecting food allergen crustacean gene |
-
2011
- 2011-11-25 CN CN 201110382763 patent/CN102363816B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101343661A (en) * | 2008-08-22 | 2009-01-14 | 华南理工大学 | Constant temperature nucleic acid amplification fast detecting reagent kit for bacillus coli O157 and use method thereof |
| CN101487057A (en) * | 2009-01-23 | 2009-07-22 | 浙江省疾病预防控制中心 | Loop-mediated isothermal amplification fast detecting reagent kit and method for O157:H7 coliform |
| CN101899501A (en) * | 2010-02-10 | 2010-12-01 | 江苏出入境检验检疫局动植物与食品检测中心 | Constant temperature amplification detection kit and method for detecting food allergen crustacean gene |
| CN101864494A (en) * | 2010-04-13 | 2010-10-20 | 上海国际旅行卫生保健中心 | Constant-temperature amplification detection kit of A(H1N1) virus and detection method thereof |
Non-Patent Citations (5)
| Title |
|---|
| RENDONG FANG 等: "Cross-Priming Amplification for Rapid Detection of Mycobacterium tuberculosis in Sputum Specimens", 《JOURNAL OF CLINICAL MICROBIOLOGY》, vol. 47, no. 3, 31 March 2009 (2009-03-31), pages 845 - 847 * |
| 周静宇 等: "交叉引物扩增技术快速检测结核分枝杆菌", 《临床检验杂志》, vol. 28, no. 1, 15 January 2010 (2010-01-15), pages 22 * |
| 宋凌浩 等: "CPA 恒温扩增技术在口岸蚊媒携带登革病毒检测中的应用", 《西南林学院学报》, vol. 30, no. 1, 15 June 2010 (2010-06-15), pages 8 - 11 * |
| 徐义刚 等: "肠出血性大肠埃希菌O157：H7环介导恒温扩增快速检测方法的建立与应用", 《中国人兽共患病学报》, vol. 26, no. 7, 15 July 2010 (2010-07-15), pages 618 - 623 * |
| 王建广 等: "大肠埃希氏菌O157:H7的HDA检测方法的建立", 《安徽农业大学学报》, vol. 38, no. 2, 23 February 2011 (2011-02-23), pages 271 - 274 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103343164A (en) * | 2013-06-17 | 2013-10-09 | 武汉轻工大学 | Multiplex PCR detection method and kit for Shiga toxin-producing Escherichia coli and application thereof |
| CN103343164B (en) * | 2013-06-17 | 2016-05-18 | 武汉轻工大学 | Shiga toxin producing escherichia coli multi-PCR detection method, kit and application |
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