CN108950037A - Dual LAMP detects Shigella and Listeria Monocytogenes primer sets, reagent, kit and method simultaneously - Google Patents
Dual LAMP detects Shigella and Listeria Monocytogenes primer sets, reagent, kit and method simultaneously Download PDFInfo
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- CN108950037A CN108950037A CN201811011429.5A CN201811011429A CN108950037A CN 108950037 A CN108950037 A CN 108950037A CN 201811011429 A CN201811011429 A CN 201811011429A CN 108950037 A CN108950037 A CN 108950037A
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Abstract
The invention discloses a kind of dual LAMP to detect Shigella and Listeria Monocytogenes primer sets simultaneously, reagent, kit and method, it passes through unique design of primers, detection while Shigella and Listeria Monocytogenes may be implemented, it is short with detection cycle, detect the advantages that speed is fast, whether the dual LAMP detection method that the present invention uses can be contains Shigella and Listeria Monocytogenes in qualitative analysis meat products, the advantages of having played LAMP detection technique, further improve the detection architecture of food-borne pathogens in meat products, increase the protection to consumer's interests.
Description
Technical field
The present invention discloses the field for being related to loop-mediated isothermal amplification technique more particularly to dual LAMP while detecting shiga
Bacterium and Listeria Monocytogenes primer sets, reagent, kit and method.
Background technique
Currently, domestic and international application technology of pathogenic bacteria in detection food mainly has traditional national standard detection method, PCR
Method, quantitative fluorescent PCR and multi-PCR detection method.
With the development of market economy, the production and consumption of meat products are always maintained at the situation of sustainable development, import and export meat
The demand of product is even more continuously increased, but Shigella and monocyte hyperplasia Listeria common in certain meat products
Bacterium uses classical culture protocols, and each pathogenic bacteria are required to individually be detected, not only workload using independent detection method
Greatly, detection cycle is long, and many factors such as profession, experience by testing staff are limited, and erroneous judgement is easy to appear.It is adopted at present
PCR method, fluorescence quantifying PCR method and multi-PCR detection method want instrument and equipment though detection efficiency can be improved
Ask higher, it is at high cost, it can not be in a wide range of interior popularization and use.
Summary of the invention
In consideration of it, the present disclosure provides a kind of dual LAMP to detect Shigella and monocyte hyperplasia Li Si simultaneously
Special Salmonella primer sets, reagent, kit and method cannot achieve the same of above-mentioned strain at least to solve previous detection method
When detect, the problems such as detection cycle is long, and False Rate is high.
One aspect of the present invention provides a kind of dual LAMP while detecting Shigella and monocyte hyperplasia Listeria
Bacterium primer sets, the primer sets include: Shigella primer sequence collection and Listeria Monocytogenes primer sequence
Column collection；
The Shigella primer sequence collection includes: outer primer ipaH-F3, outer primer ipaH-B3, inner primer ipaH-FIP
And inner primer ipaH-BIP；
The outer primer F3 has SEQ No.1 sequence；
The outer primer B3 has SEQ No.2 sequence；
The inner primer FIP has SEQ No.3 sequence；
The inner primer BIP has SEQ No.4 sequence；
The Listeria Monocytogenes primer sequence collection include: outer primer hlyA-F3, outer primer hlyA-B3,
Inner primer hlyA-FIP and inner primer hlyA-BIP；
The outer primer F3 has SEQ No.5 sequence；
The outer primer B3 has SEQ No.6 sequence；
The inner primer FIP has SEQ No.7 sequence；
The inner primer BIP has SEQ No.8 sequence.
Shigella is detected simultaneously the present invention also provides a kind of dual LAMP and Listeria Monocytogenes are used
Reagent, the reagent contain above-mentioned primer sets.
The present invention additionally provides a kind of dual LAMP simultaneously while detecting Shigella and monocyte hyperplasia Listeria
Bacterium kit, the kit contain above-mentioned primer sets or above-mentioned reagent.
It is preferred that the dual LAMP detects Shigella and Listeria Monocytogenes kit simultaneously, also
Contain reaction buffer, Bst archaeal dna polymerase, deionized water, positive reference substance and negative controls.
Further preferably, the positive reference substance is Shigella DNA template and Listeria Monocytogenes
DNA template.
Further preferably, the negative controls are non-Shigella and Listeria Monocytogenes because of a group DNA.
Another aspect of the present invention additionally provides a kind of dual LAMP while detecting Shigella and monocyte hyperplasia Li Si
The method of special Salmonella, the method are detected using mentioned reagent box, are included the following steps:
1) genome of sample to be tested DNA is extracted；
2) using extracted DNA as template, primer sets, reaction buffer, Bst archaeal dna polymerase and deionization is added
Water carries out LAMP reaction, and records amplification, wherein reaction temperature is 63 DEG C, reaction time 60min；
3) positive control and negative control are set, wherein positive control is with Shigella and monocyte hyperplasia Liszt
Salmonella genomic DNA is template, according to condition described in step 2), carries out LAMP detection, records amplification, negative control is with non-
Shigella and Listeria Monocytogenes, according to condition described in step 2), carry out LAMP inspection because group DNA is template
It surveys, records amplification；
4) if the amplification in step 2) is shown as without amplification phenomenon, and the amplification of positive control shows to have and expand now
As, and the amplification of negative control is shown without amplification phenomenon, then determines that Shigella and monokaryon are not detected in sample to be tested
Monocytogenes；If the amplification in step 2) is shown with amplification phenomenon, and the amplification of positive control
It is aobvious to have amplification phenomenon, and the amplification of negative control is shown without amplification phenomenon, then determines to detect shiga in sample to be tested
Bacterium and Listeria Monocytogenes.
The same of Shigella and Listeria Monocytogenes may be implemented by unique design of primers in the present invention
When detect, detection cycle is short, detection speed it is fast.Loop-mediated isothermal amplification technique (the Loop-mediated that the present invention uses
Isothermal amplification, LAMP) it is a kind of novel constant temperature nucleic acid amplification technology, since its unique nucleic acid expands
Increasing mechanism, product can show LAMP on agarose gel electrophoresis and react typical ladder-like band, meanwhile, nucleic acid is largely given birth to
Cheng Shi, the Mg from the pyrophosphate ion and reaction system being precipitated in dNTPs2+In conjunction with generating macroscopic amplified reaction
By-product --- white magnesium pyrophosphate precipitating, therefore LAMP amplification not only view mode multiplicity, and result identification simplicity, accidentally
Sentence that rate is low, is very suitable to high-throughput quick detection.Have the advantages that 1, detection cycle is short, as pathogenic bacteria prescreening method
The time can be shortened, reduce cost, speed passenger flow speed, improves working efficiency；2, detection device is small in size, is convenient for carrying；3,
Sensitivity is higher, expands simultaneously.
Detailed description of the invention
The drawings herein are incorporated into the specification and forms part of this specification, and shows and meets implementation of the invention
Example, and be used to explain the principle of the present invention together with specification.
Fig. 1 discloses embodiment for the present invention and detects Shigella and Listeria Monocytogenes in chicken meat sample
LAMP amplification figure；
Fig. 2 is LAMP amplification figure when detecting in the embodiment of the present invention about sensitivity；
LAMP amplification figure when Fig. 3 is in the embodiment of the present invention about specific detection；
Fig. 4 is LAMP amplification figure when detecting in the embodiment of the present invention about repeatability.
Specific embodiment
The protection model that the present invention is further expalined with specific case below, but is not intended to restrict the invention
It encloses.
Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art,
Product used is commercially available.
Detection process of the invention is described in further detail below with reference to specific embodiment, specific as follows:
1, design of primers
Select the ipaH gene (GenBank:M32063.1) and Listeria Monocytogenes of Shigella
HlyA gene (GenBank:DQ812517.1) is used as target sequence, is distinguished by Primer Explore V4 Photographing On-line software
Shigella primer sequence collection and Listeria Monocytogenes primer sequence collection are designed, table specific as follows:
2, the extraction and purification of sample DNA
The nutrient broth of bacterium will be increased, 1mL is directly taken to be added in 1.5mL sterile centrifugation tube, 8000rpm is centrifuged 4min, as far as possible
It inhales and abandons supernatant；750 μ L DNA extracting solutions are added, boiling water bath 5min adds phenol: 700 μ L of chloroform (1:1, volume ratio), oscillation are mixed
Even, 13000rpm is centrifuged 5min, pipettes supernatant, and the isopropanol precipitating nucleic acid of 0.6 times of volume, 13000rpm centrifugation is added
10min is discarded supernatant, and is precipitated once with 70% ethanol washing of 1mL, and 13000rpm is centrifuged 10min, discards supernatant, precipitating is dissolved in
In 20 μ L nucleic acid lysates.Be stored in -20 DEG C it is spare.- 70 DEG C can long-term preservation.Equivalent commercialized DNA can also be used to mention
It takes kit and illustrates that extraction prepares template DNA by it.
3, LAMP reaction amplification system is established
LAMP reaction system is 25 μ L: 12.5 μ L of reaction buffer, Shigella and Listeria Monocytogenes
Each 0.5 μ L of inner primer FIP (40 μm of ol/L), each 0.5 μ L of inner primer BIP (40 μm of ol/L), each 0.5 μ L of outer primer F3 (5 μm of ol/
L), each 0.5 μ L of outer primer B3 (5 μm of ol/L), 1 μ L of Bst archaeal dna polymerase, 5.5 μ L of deionized water, Shigella and monocyte
Each 1 μ L of monocytogenes DNA template, negative control are 2 μ L deionized waters.Use DNA amplification kit
(LAMP method) carries out LAMP amplification test on transmissometer LA-320, and 63 DEG C of reaction 60min record amplification, and reaction terminates
80 DEG C of inactivation 5min afterwards.
4, quality control index is determined
Positive control is using Shigella and Listeria Monocytogenes genomic DNA as template, according to 3 items
Part carries out LAMP detection, records amplification；Negative control is with non-Shigella and Listeria Monocytogenes because of group
DNA is template, according to 3 conditions, carries out LAMP detection, records amplification.
5, result judgement
If the amplification of sample to be tested is shown as without amplification phenomenon, and the amplification of positive control shows to have and expand now
As, and the amplification of negative control is shown without amplification phenomenon, then determines that Shigella and monokaryon are not detected in sample to be tested
Monocytogenes；If the amplification of sample to be tested is shown with amplification phenomenon, and the amplification of positive control
It is aobvious to have amplification phenomenon, and the amplification of negative control is shown without amplification phenomenon, then determines to detect shiga in sample to be tested
Bacterium and Listeria Monocytogenes.
6, pattern detection and result verification
Shigella (ATCC 12022) and Listeria Monocytogenes (CMCC 54002) are inoculated in respectively
Culture solution is centrifuged by nutrient broth culture solution, 36 DEG C of overnight incubations respectively, after discarding supernatant liquid, be added physiological saline repeatedly from
The heart washs 3 times, adds physiological saline and suspends again, and adjust bacteria suspension concentration, is uniformly mixed in chicken processed sample, makes sample
Contaminant capacity in product reaches 103/mL(g).Analog sample is detected in two ways respectively.Method one: 25g chicken is taken to add respectively
Work sample is added in the BPW enrichment liquid of 225mL, after 36 DEG C of culture 12h, using DNA of bacteria in kit extraction culture solution, and with
This is template, is detected using dual LAMP method；Method two: by chicken processed sample according to the method for concerned countries standard
Qualitative detection is carried out, and two groups of testing results are compared.
According to above-mentioned detection process, after chicken processed goods is extracted DNA, LAMP detection is carried out, wherein corresponding LAMP expands
Increase curve and see Fig. 1, show amplification phenomenon, and positive control amplification it is aobvious have an amplification phenomenon, and the amplification of negative control
As the result is shown without amplification phenomenon, Fig. 1 is specifically also seen, therefore, determine chicken processing variety detection Shigella and monocyte
Monocytogenes.
Chicken processed sample is detected using GB 4789.5-2012 and GB 4789.30-2016 national standard method respectively simultaneously,
As a result Shigella and Listeria Monocytogenes are detected, it is consistent with dual LAMP detection method result.
7, sensitivity detects
By the hybrid dna template of Shigella and Listeria Monocytogenes and carry out 10 times of gradient dilutions.For
The sensitivity of the dual LAMP detection method of evaluation carries out dual LAMP amplified reaction to Mixed Microbes DNA template.As the result is shown:
Dual LAMP detection method sensitivity is higher, can reach 7.27pg/uL, be specifically shown in Fig. 2.
8, specific detection
All bacterial genomes of extraction are detected using reaction system in 3, examine the special of dual LAMP method
Property.By 23 plants of strains testeds, (including 4 plants of Shigellas, 1 plant of Listeria Monocytogenes are related to Si Shi Listeria
18 plants of non-targeted bacterial strains such as bacterium) culture solution through bacterium extract be made DNA profiling, carry out dual LAMP amplification, amplification table
Bright: except the positive is presented in Shigella and Listeria Monocytogenes amplification, remaining bacterial strain is feminine gender, it was demonstrated that should
Dual LAMP detection method specificity is good, is specifically shown in Fig. 3.
9, repeatability detection
Chicken meat sample in above-described embodiment is repeated 6 times detection, result is the positive, referring specifically to Fig. 4.
Above-described embodiment use dual LAMP detection method can in qualitative analysis meat products whether containing Shigella and
Listeria Monocytogenes, the advantages of having played LAMP detection technique, food-borne cause in further perfect meat products
The detection architecture of germ is increased to the protections of consumer's interests, but because dual LAMP detection simultaneously amplification judging whether
Detection, cannot distinguish two kinds of pathogenic bacteria simultaneously, need if any the positive with single LAMP or PCR method difference amplification judging.
Those skilled in the art after considering the specification and implementing the invention disclosed here, will readily occur to of the invention its
Its embodiment.This application is intended to cover any variations, uses, or adaptations of the invention, these modifications, purposes or
Person's adaptive change follows general principle of the invention and including the undocumented common knowledge in the art of the present invention
Or conventional techniques.The description and examples are only to be considered as illustrative, and true scope and spirit of the invention are by following
Claim is pointed out.
It should be understood that the invention is not limited to the content being described above, can without departing from the scope into
Row various modifications and change.The scope of the present invention is limited only by the attached claims.
Sequence table
<110>Shenyang Entry-Exit Inspection and Quarantine Bureau inspection and quarantine complex art center
<120>dual LAMP detects Shigella and Listeria Monocytogenes primer sets, reagent, reagent simultaneously
Box and method
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Claims (7)
1. a kind of dual LAMP detects Shigella and Listeria Monocytogenes primer sets simultaneously, feature exists
In the primer sets include: Shigella primer sequence collection and Listeria Monocytogenes primer sequence collection；
The Shigella primer sequence collection include: outer primer ipaH-F3, outer primer ipaH-B3, inner primer ipaH-FIP and
Inner primer ipaH-BIP；
The outer primer F3 has SEQ No.1 sequence；
The outer primer B3 has SEQ No.2 sequence；
The inner primer FIP has SEQ No.3 sequence；
The inner primer BIP has SEQ No.4 sequence；
The Listeria Monocytogenes primer sequence collection includes: outer primer hlyA-F3, outer primer hlyA-B3, interior draws
Object hlyA-FIP and inner primer hlyA-BIP；
The outer primer F3 has SEQ No.5 sequence；
The outer primer B3 has SEQ No.6 sequence；
The inner primer FIP has SEQ No.7 sequence；
The inner primer BIP has SEQ No.8 sequence.
2. a kind of dual LAMP detects Shigella and Listeria Monocytogenes reagent simultaneously, which is characterized in that
The reagent contains primer sets described in claim 1.
3. a kind of dual LAMP detects Shigella and Listeria Monocytogenes kit simultaneously, feature exists
In the kit contains primer sets described in claim 1 or reagent as claimed in claim 2.
4. dual LAMP detects Shigella and Listeria Monocytogenes reagent simultaneously according to claim 3
Box, which is characterized in that the kit also contain reaction buffer, Bst archaeal dna polymerase, deionized water, positive reference substance with
And negative controls.
5. dual LAMP detects Shigella and Listeria Monocytogenes reagent simultaneously according to claim 4
Box, which is characterized in that the positive reference substance is Shigella DNA template and Listeria Monocytogenes DNA mould
Version.
6. dual LAMP detects Shigella and Listeria Monocytogenes reagent simultaneously according to claim 4
Box, which is characterized in that the negative controls are non-Shigella and Listeria Monocytogenes because of a group DNA.
7. a kind of method that dual LAMP detects Shigella and Listeria Monocytogenes simultaneously, which is characterized in that
Using kit described in claim 4, include the following steps:
1) genome of sample to be tested DNA is extracted；
2) using extracted DNA as template, be added primer sets, reaction buffer, Bst archaeal dna polymerase and deionized water into
Row LAMP reaction, and record amplification, wherein reaction temperature is 63 DEG C, reaction time 60min；
3) positive control and negative control are set, wherein positive control is with Shigella and Listeria Monocytogenes
Genomic DNA is template, according to condition described in step 2), carries out LAMP detection, records amplification, negative control is congratulated with non-will
Salmonella and Listeria Monocytogenes, according to condition described in step 2), carry out LAMP detection, note because group DNA is template
Record amplification；
If the amplification 4) in step 2) is shown as without amplification phenomenon, and positive control amplification it is aobvious have amplification phenomenon,
And the amplification of negative control is shown without amplification phenomenon, then determines that Shigella and monocyte are not detected in sample to be tested
Monocytogenes；If the amplification in step 2) is shown with amplification phenomenon, and the amplification of positive control aobvious has
Expand phenomenon, and the amplification of negative control show without amplification phenomenon, then determine in sample to be tested detect Shigella with
Listeria Monocytogenes.
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Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113475A (en) * | 2007-05-30 | 2008-01-30 | 中国疾病预防控制中心传染病预防控制所 | Primer for detecting pathogenic microorganism and multiple PCR using the same |
| CN101434992A (en) * | 2008-09-28 | 2009-05-20 | 中国计量学院 | Fluorescent quantitative PCR detecting method for Listeria monocytogenes and Shigella |
| CN101603070A (en) * | 2008-06-12 | 2009-12-16 | 陈福生 | A kind of method of rapid detection shigella |
| CN101603071A (en) * | 2008-06-12 | 2009-12-16 | 陈福生 | The method of a kind of while rapid detection salmonella and shigella |
| CN102220427A (en) * | 2011-05-10 | 2011-10-19 | 浙江省质量技术监督检测研究院 | Multiple PCR (Polymerase Chain Reaction) detection method for four food-borne pathogens, detection primer set and kit |
| CN102586439A (en) * | 2012-02-28 | 2012-07-18 | 盛司潼 | Method for simultaneously and quickly detecting multiple nucleic acids |
| CN102703588A (en) * | 2011-08-09 | 2012-10-03 | 中国人民解放军后勤工程学院 | Multiplex PCR-based synchronous and rapid method for detecting 13 pathogenic microorganisms in water |
| CN103898208A (en) * | 2013-11-27 | 2014-07-02 | 中华人民共和国上海出入境检验检疫局 | Quick high-throughput intestines source pathogenic bacterium detection method |
| CN104313173A (en) * | 2014-11-11 | 2015-01-28 | 舟山市质量技术监督检测研究院 | LAMP method for detecting real-time turbidity of Listeria monocytogenes |
| CN106544424A (en) * | 2016-10-31 | 2017-03-29 | 中国疾病预防控制中心传染病预防控制所 | The constant-temperature amplifications that intersect combine the method that gold nano bio-sensing detects shigella dysenteriae more |
| CN106811535A (en) * | 2017-03-08 | 2017-06-09 | 青岛捷安信检验技术服务有限公司 | It is a kind of at the same detect five kinds of primer combination of probe things and multiple real time fluorescence PCR method of pathogenic bacteria |
| CN107022644A (en) * | 2017-06-14 | 2017-08-08 | 山东省农业科学院农业质量标准与检测技术研究所 | Six kinds of multiple LAMP detection primers of food-borne pathogens, detection kit and detection method in fruits and vegetables |
| CN108220464A (en) * | 2018-04-08 | 2018-06-29 | 陈思 | A kind of 16 kinds of food-borne pathogens detection kits |
-
2018
- 2018-08-31 CN CN201811011429.5A patent/CN108950037A/en not_active Withdrawn
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113475A (en) * | 2007-05-30 | 2008-01-30 | 中国疾病预防控制中心传染病预防控制所 | Primer for detecting pathogenic microorganism and multiple PCR using the same |
| CN101603070A (en) * | 2008-06-12 | 2009-12-16 | 陈福生 | A kind of method of rapid detection shigella |
| CN101603071A (en) * | 2008-06-12 | 2009-12-16 | 陈福生 | The method of a kind of while rapid detection salmonella and shigella |
| CN101434992A (en) * | 2008-09-28 | 2009-05-20 | 中国计量学院 | Fluorescent quantitative PCR detecting method for Listeria monocytogenes and Shigella |
| CN102220427A (en) * | 2011-05-10 | 2011-10-19 | 浙江省质量技术监督检测研究院 | Multiple PCR (Polymerase Chain Reaction) detection method for four food-borne pathogens, detection primer set and kit |
| CN102703588A (en) * | 2011-08-09 | 2012-10-03 | 中国人民解放军后勤工程学院 | Multiplex PCR-based synchronous and rapid method for detecting 13 pathogenic microorganisms in water |
| CN102586439A (en) * | 2012-02-28 | 2012-07-18 | 盛司潼 | Method for simultaneously and quickly detecting multiple nucleic acids |
| CN103898208A (en) * | 2013-11-27 | 2014-07-02 | 中华人民共和国上海出入境检验检疫局 | Quick high-throughput intestines source pathogenic bacterium detection method |
| CN104313173A (en) * | 2014-11-11 | 2015-01-28 | 舟山市质量技术监督检测研究院 | LAMP method for detecting real-time turbidity of Listeria monocytogenes |
| CN106544424A (en) * | 2016-10-31 | 2017-03-29 | 中国疾病预防控制中心传染病预防控制所 | The constant-temperature amplifications that intersect combine the method that gold nano bio-sensing detects shigella dysenteriae more |
| CN106811535A (en) * | 2017-03-08 | 2017-06-09 | 青岛捷安信检验技术服务有限公司 | It is a kind of at the same detect five kinds of primer combination of probe things and multiple real time fluorescence PCR method of pathogenic bacteria |
| CN107022644A (en) * | 2017-06-14 | 2017-08-08 | 山东省农业科学院农业质量标准与检测技术研究所 | Six kinds of multiple LAMP detection primers of food-borne pathogens, detection kit and detection method in fruits and vegetables |
| CN108220464A (en) * | 2018-04-08 | 2018-06-29 | 陈思 | A kind of 16 kinds of food-borne pathogens detection kits |
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Application publication date: 20181207 |