CN106011154A - Molecular detection kit for klebsiella pneumoniae causing hepatic abscess and application thereof - Google Patents
Molecular detection kit for klebsiella pneumoniae causing hepatic abscess and application thereof Download PDFInfo
- Publication number
- CN106011154A CN106011154A CN201610669762.XA CN201610669762A CN106011154A CN 106011154 A CN106011154 A CN 106011154A CN 201610669762 A CN201610669762 A CN 201610669762A CN 106011154 A CN106011154 A CN 106011154A
- Authority
- CN
- China
- Prior art keywords
- klebsiella pneumoniae
- liver abscess
- luxr
- pago
- abscess
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a molecular detection kit for klebsiella pneumoniae causing hepatic abscess and an application thereof. The invention provides gene specific sequences pagO and luxR of klebsiella pneumoniae causing hepatic abscess, two genes are specially existed in the klebsiella pneumoniae causing hepatic abscess, and can be taken as a diagnostic marker of klebsiella pneumoniae causing hepatic abscess. The primers of design of two genes can be used for preparing the molecular detection kit for klebsiella pneumoniae causing hepatic abscess, the kit can be used for diagnosis or early diagnosis of the molecular detection kit for klebsiella pneumoniae causing hepatic abscess, and has great commercialized exploitation value.
Description
Technical field
The present invention relates to medical diagnosis on disease field, be specifically related to a kind of cause liver abscess Klebsiella Pneumoniae molecular detection kit
And application.
Background technology
Klebsiella Pneumoniae (K.pneumoniae subsp.pneumonia), belongs to enterobacteriaceae lactobacteriaceae.Often parasitize
Human body skin, nasopharynx part and intestinal etc., for conditioned pathogen.Clinically, pneumonia and urinary tract infection etc. can be caused
(Podschun R,Ullmann U.Klebsiella spp.as nosocomial pathogens:epidemiology,
taxonomy,typing methods,and pathogenicity factors.Clinical microbiology
reviews.1998；11(4):589-603).Within 1980, Taiwan Liu et al. reported first 7 examples are by high poison to nineteen ninety
Serious liver abscess companion endophthalmitis case (Liu Y, Cheng D, the Lin C.Klebsiella that power Klebsiella Pneumoniae causes
pneumoniae liver abscess associated with septic endophthalmitis.Arch Intern
Med 1986；146:1913–16).Hereafter, Klebsiella Pneumoniae liver abscess starts Global prevalence, particularly in Asia.With commonly
Klebsiella Pneumoniae compare, cause liver abscess Klebsiella Pneumoniae aggressivity is strong, fatality rate is high.Except causing liver abscess, cause liver
Abscess Klebsiella Pneumoniae can also cause serious meningitis, endophthalmitis and pulmonary abscess etc. (Siu LK, Yeh KM, Lin JC,
Fung CP,Chang FY.Klebsiella pneumoniae liver abscess:a new invasive
syndrome.The Lancet Infectious diseases.2012；12(11):881-7).Lung for the highest virulence
Scorching Klebsiella infection, diagnoses as early as possible, treats in time, and the generation to reduction severe complication improves prognosis and has important facing
Bed is worth.But, currently without a kind of fast and convenient method for causing the Testing and appraisal of liver abscess Klebsiella Pneumoniae.
Along with the increase of Klebsiella Pneumoniae liver abscess Case report and going deep into of cause liver abscess Klebsiella Pneumoniae research,
Several phenotypes relevant to causing liver abscess Klebsiella Pneumoniae are suggested, in succession including bacterium colony high viscosity phenotype and capsular polysaccharide blood
Clear type K1 type/K2 type etc..Klebsiella Pneumoniae can secrete polysaccharose substance in cell surface, so that bacterium colony produces high viscosity table
Type, in order to resist the phagocytosis of macrophage and neutrophilic granulocyte.But study discovery recently, only 50% less than cause liver abscess lung
Scorching klebsiella shows as bacterium colony high viscosity (Qu TT, Zhou JC, Jiang Y, Shi KR, Li B, Shen P, et
al.Clinical and microbiological characteristics of Klebsiella pneumoniae
liver abscess in East China.BMC infectious diseases.2015；15:161).Capsular polysaccharide serotypes
Antigen K1 type and K2 type are the predominant serotypes causing liver abscess Klebsiella Pneumoniae, but still have the cause liver abscess pneumonia of more than 20%
Klebsiella is not belonging to K1/K2 serotype.Additionally, serotype K1 type or K2 type are also found to be present in pneumonia klebsiella
(Sun Y,Wu H,Shen D.Clinical and Molecular Analysis of Klebsiella pneumoniae
Causing Liver Abscess in China.Journal of molecular microbiology and
biotechnology.2016；26(4):245-51).Therefore, in order to cause liver more rapid effective qualification of molecular level
Abscess Klebsiella Pneumoniae, we are by causing liver abscess Klebsiella Pneumoniae and the genome ratio of pneumonia klebsiella
Relatively analyze, find the specific gene sequences causing liver abscess Klebsiella Pneumoniae.Utilize these specific sequences, by simply
PCR method, can identify cause liver abscess Klebsiella Pneumoniae rapidly, for by the microbial infection of this cause of disease in time diagnosis and
Treat, patient's prognosis offer basis is provided.
Summary of the invention
Object of the present invention is to provide cause liver abscess Klebsiella Pneumoniae gene specific sequence pagO, described base
Because specific sequence pagO is shown in SEQ ID NO.1.
Further object is that and provide based on causing liver abscess Klebsiella Pneumoniae gene specific sequence
The primer of pagO design, primer is: pagO_F:TGCTCTTGAAACTATCCCTCC, pagO_R:GGCAATAACTCCCGTCCA.
Cause liver abscess Klebsiella Pneumoniae gene specific sequence luxR, institute are it is also an object of the present invention to provide
The gene specific sequence luxR stated is shown in SEQ ID NO.2.
It is also an object of the present invention to provide based on causing liver abscess Klebsiella Pneumoniae gene specific sequence
The primer of luxR design, primer is: luxR_F:CTTTGCCGGCATGGAACATA, luxR_R:
TGAGCCAAATGTATGCCAAGGA。
It is also an object of the present invention to provide cause liver abscess Klebsiella Pneumoniae gene specific sequence pagO or
The primer based on pagO design application in preparation causes liver abscess Klebsiella Pneumoniae detection kit, including for preparing lung
Application in scorching klebsiella liver abscess early diagnosis kit.
It is also an object of the present invention to provide cause liver abscess Klebsiella Pneumoniae gene specific sequence luxR or
The primer based on luxR design application in preparation causes liver abscess Klebsiella Pneumoniae detection kit, including for preparing lung
Application in scorching klebsiella liver abscess early diagnosis kit.
Final object of the present invention there are provided a kind of cause liver abscess Klebsiella Pneumoniae molecular detection kit,
Described test kit includes primer:
PagO_F:TGCTCTTGAAACTATCCCTCC
PagO_R:GGCAATAACTCCCGTCCA
LuxR_F:CTTTGCCGGCATGGAACATA
LuxR_R:TGAGCCAAATGTATGCCAAGGA.
To achieve these goals, the present invention uses techniques below measure:
Causing liver abscess Klebsiella Pneumoniae gene specific sequence pagO, described gene specific sequence pagO is SEQ
Shown in ID NO.1.
Primer based on pagO sequential design is protection scope of the present invention, it is preferred that the primer of pagO is: pagO_
F:TGCTCTTGAAACTATCCCTCC；PagO_R:GGCAATAACTCCCGTCCA.
Causing liver abscess Klebsiella Pneumoniae gene specific sequence luxR, described gene specific sequence luxR is SEQ
Shown in ID NO.2.
Primer based on luxR sequential design is protection scope of the present invention, it is preferred that the primer of luxR is: luxR's
Primer is: luxR_F:CTTTGCCGGCATGGAACATA；LuxR_R:TGAGCCAAATGTATGCCAAGGA.
Protection scope of the present invention also include cause liver abscess Klebsiella Pneumoniae gene specific sequence pagO or based on
The application in preparation causes liver abscess Klebsiella Pneumoniae detection kit of the primer of pagO design, including preparing kerekou pneumonia primary
Bacterium liver abscess early diagnosis kit.
Protection scope of the present invention also include cause liver abscess Klebsiella Pneumoniae gene specific sequence luxR or based on
The application in preparation causes liver abscess Klebsiella Pneumoniae detection kit of the primer of luxR design, including preparing kerekou pneumonia primary
Bacterium liver abscess early diagnosis kit.
A kind of cause liver abscess Klebsiella Pneumoniae detection kit, including:
PagO_F:TGCTCTTGAAACTATCCCTCC
PagO_R:GGCAATAACTCCCGTCCA
LuxR_F:CTTTGCCGGCATGGAACATA
LuxR_R:TGAGCCAAATGTATGCCAAGGA.
Compared with prior art, the invention have the characteristics that:
There is no well for causing the molecular marker that liver abscess Klebsiella Pneumoniae is identified the most at present, the present invention is by surveying
Sequence 2 strain is isolatable from cause liver abscess Klebsiella Pneumoniae and the 1 strain non-cause liver abscess Klebsiella Pneumoniae of China's Mainland, and associating is online
45 strains announced cause liver abscess Klebsiella Pneumoniae and the full-length genome data of 103 strains non-cause liver abscess Klebsiella Pneumoniae, logical
Cross comparison analysis, obtained for causing two sections of sequences that liver abscess Klebsiella Pneumoniae camber is special.
2. the sequence that 2 sections of cause liver abscess Klebsiella Pneumoniaes of present invention offer are special, can cause liver abscess as preparation
Klebsiella Pneumoniae detection kit
3. the sequence that 2 sections of cause liver abscess Klebsiella Pneumoniaes of present invention offer are special, can use as molecular marker
Early diagnosis in Klebsiella Pneumoniae liver abscess.Particularly in the Bacteria culturing result of other body fluid source specimen such as blood
Before going out report, utilize blood samples of patients specimen or liver vomica pus, use the detection of quick PCR method to cause liver abscess kerekou pneumonia
Primary bacterium specific gene pagO is or/and the presence or absence of luxR, in conjunction with patients with clinical manifestations and imaging data, and can be quickly to pneumonia
Klebsiella liver abscess makes diagnosis, improves the treatment success rate of Klebsiella Pneumoniae liver abscess.
Detailed description of the invention
The invention will be further described for specific embodiment.Technical scheme of the present invention, if not otherwise specified, is this
The conventional scheme in field.
Embodiment 1:
The exploitation of specific sequence and checking:
The screening process of specific gene sequences combines second filial generation sequencing technologies (Next-Generation
Sequencing, NGS), comparative genomics (Comparative Genomics) and bioinformatics
(Bioinformatics) method, and use the statistical method of science, to sequence in multiple Klebsiella Pneumoniae genomes
The specificity of distribution is tested, and at the cause liver abscess Klebsiella Pneumoniae separated and non-cause liver abscess Klebsiella Pneumoniae
In carry out PCR checking, to determine that sequence is at the specificity caused in liver abscess Klebsiella Pneumoniae.Specific implementation process is as follows:
Choose 2 strains and be isolatable from cause liver abscess Klebsiella Pneumoniae and the 1 strain non-cause liver abscess kerekou pneumonia primary of China's Mainland
Bacterium is used for genome sequencing, and sequencing result SPAdes (v3.5) software splices, and enters with PATRIC on-line analysis platform
Row predictive genes and functional annotation.Sequencing result associating US National Bioinformatics Institute (the National Center obtained
For Bi otechnology Information, NCBI) 45 strains announced cause liver abscess Klebsiella Pneumoniae and the non-causes of 103 strains
The full-length genome data of liver abscess Klebsiella Pneumoniae, are analyzed by comparison, and screening exists only in cause liver abscess kerekou pneumonia primary
Gene in bacterium genome, has obtained two sections and has caused sequence pagO (the SEQ ID that liver abscess Klebsiella Pneumoniae camber is special
Shown in NO.1) and luxR (shown in SEQ ID NO.2) based on these two sections of special primers, exploitation obtain a set of for
The primer of qualification cause liver abscess Klebsiella Pneumoniae:
The primer of pagO is:
PagO_F:TGCTCTTGAAACTATCCCTCC
PagO_R:GGCAATAACTCCCGTCCA
The primer of luxR is:
LuxR_F:CTTTGCCGGCATGGAACATA
LuxR_R:TGAGCCAAATGTATGCCAAGGA.
Embodiment 2:
Gene specific is verified:
PagO and luxR causes the checking in Clinical isolation of the liver abscess Klebsiella Pneumoniae specific gene, and it is concrete
Step is:
1. select the cause liver abscess that is clinically separated and non-cause liver abscess Klebsiella Pneumoniae bacterial strain, be used for detecting pagO (SEQ
ID N is O.1 shown) and the luxR (shown in SEQ ID NO.2) specificity in causing liver abscess Klebsiella Pneumoniae.Select liver pus
The 40 strain Klebsiella Pneumoniaes separated in swollen patient's vomica, as detection group (causing liver abscess group), select negated Klebsiella Pneumoniae else
Klebsiella Pneumoniae 36 strain separated in liver abscess patient is as negative control group (non-cause liver abscess group).Also have chosen simultaneously and face
Other pathogenic bacterium common (other pathogenic bacterium groups) separated on bed are to verify that these factors are causing liver abscess kerekou pneumonia further
Specificity in primary bacterium: include staphylococcus aureus, escherichia coli, Acinetobacter bauamnnii, enterococcus faecalis, enterococcus faecalis.
2. the design of 2 predicted gene specific primers: for pagO and luxR specific sequence design primer such as
Under:
The primer of pagO is:
PagO_F:TGCTCTTGAAACTATCCCTCC
PagO_R:GGCAATAACTCCCGTCCA
The primer of luxR is:
LuxR_F:CTTTGCCGGCATGGAACATA
LuxR_R:TGAGCCAAATGTATGCCAAGGA
3. the extraction of bacterial genomes: the 76 strain Klebsiella Pneumoniaes chosen are extracted DNA in a conventional manner.
4.PCR detection pagO and luxR gene (causes liver abscess group, non-cause liver abscess group and other pathogenic bacterium three groups of bacterium
Group) in existence situation: PCR reaction use Premix TaqVersion 2.0 (TaKaRa, Code:D331A) test kit.Root
Illustrating according to test kit, the PCR reaction system of setting is 25ul, wherein Premix Taq 12.5ul, genomic DNA template 1ul
(about 100ng), upstream and downstream primer 0.5ul (10uM), add deionized water 10.5ul and be supplemented to cumulative volume 25ul.
PCR condition is: 94 DEG C of degeneration 5min；95 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 30s, 30 circulations；72℃7min；Reaction
The 5ul product agarose gel electrophoresis for DNA is respectively taken, running gel ethidium bromide (Ethi dium after end
Bromid) after dyeing 5min, photographic analysis on U.S. Bole (Bio-Rad) gel image analyser.
The results are shown in Table 1, as shown in table 1, pagO and luxR detection positive rate in causing liver abscess Klebsiella Pneumoniae divides
It is not 100%, and is all 2.78% in the Klebsiella Pneumoniae of non-cause liver abscess.In other pathogenic bacterium groups, including golden yellow
Color staphylococcus, escherichia coli, Acinetobacter bauamnnii, enterococcus faecalis, enterococcus faecalis are feminine gender.
Result above may certify that, pagO or luxR has the highest specificity in causing liver abscess Klebsiella Pneumoniae,
Can respectively or be combined as the molecular marked compound of diagnosis of pneumonia klebsiella liver abscess.
The result of table 1 pagO and luxR PCR detection in clinical strains
Embodiment 3:
PagO and luxR sequence or primer based on its sequential design are preparing Klebsiella Pneumoniae liver abscess diagnostic reagent
Application in box:
This experimental design thinking is as follows: the hemoculture sample that clinic obtains is divided into blood culture with positive bacteria group and Healthy People negative
Group, wherein blood culture with positive bacteria group turns out the sample of antibacterial in referring to all blood, is not limited solely in blood turn out kerekou pneumonia
Primary bacterium, including the sample turning out other pathogenic bacterium.With special in PCR detection positive blood culture sample and negative hemoculture sample
The existence situation of sex factor (i.e. pagO and luxR), finally PCR result and patient's final clinical diagnosis correspondence analysis, comes with this
Judge method that the present invention the provides recall rate to Klebsiella Pneumoniae liver abscess.
1. the collection of clinical hemoculture sample: collect 2 groups of hemoculture samples, blood culture with positive bacteria group 69 bottles, healthy human blood altogether
Cultivate feminine gender group 40 bottles.
2.PCR detects pagO and luxR existence situation in hemoculture sample:
The preparation of 2.1 templates: aseptically, the blood in extraction 3ml culture bottle, 8000rpm, centrifugal 2min.Abandon
Supernatant, the piping and druming of precipitation 200ul distilled water is gone to be mixed into suspension.Suspension boils 10min at 100 DEG C, then with 13000rpm, from
Heart 5min, the template that supernatant detects as PCR.
2.2PCR detection pagO and the luxR gene order label and combinations thereof existence feelings in two groups of hemoculture samples
Condition: PCR reaction uses Premix TaqVersion2.0 (TaKaRa, Code:D331A) test kit.Illustrate according to test kit, if
The PCR reaction system put is 25ul, wherein Premix Taq 12.5ul, genomic DNA template 2ul, upstream and downstream primer 0.5ul
(10uM), add deionized water 10.5ul and be supplemented to cumulative volume 25ul.PCR reaction condition is: 94 DEG C of degeneration 5min；95 DEG C of 30s, 55
DEG C 30s, 72 DEG C of 30s, 30 circulations；72℃7min；Reaction respectively takes the 5ul product agarose gel for DNA after terminating
Electrophoresis, running gel is with after ethidium bromide (Ethidium bromid) dyeing 5min, at U.S. Bole (Bio-Rad) gel imaging
Photographic analysis on analyser.
The primer of pagO is:
PagO_F:TGCTCTTGAAACTATCCCTCC
PagO_R:GGCAATAACTCCCGTCCA
The primer of luxR is:
LuxR_F:CTTTGCCGGCATGGAACATA
LuxR_R:TGAGCCAAATGTATGCCAAGGA
In blood culture with positive bacteria group, PCR testing result and final patient's confirmed result are shown in Table 2.Regulation, in pagO and luxR only
Wanting an amplification is that the positive i.e. this sample is for positive.Healthy People hemoculture feminine gender group expands without any band, is feminine gender,
Therefore result is not shown in table 2 and table 3." confirmed result " hurdle in table 2 "+" last diagnostic that represents patient is kerekou pneumonia primary
Bacterium liver abscess, "-" represents that patient gets rid of the diagnosis of Klebsiella Pneumoniae liver abscess, for other diseases.Atopen hurdle (i.e. " pa
GO ", " luxR ", " pagO+luxR ") in "+" represent that >=1 label amplification be the positive, "-" represents the detected factor
Complete negative.
The table 2 blood culture with positive bacteria final confirmed result of group patient and PCR amplification
Wherein in table 2, atopen PCR result in the hemoculture sample of No. 10 cases is all positive, but at the beginning
No. 10 cases clinically and have not been diagnosed as Klebsiella Pneumoniae liver abscess, and applicant suspects that the factor occurs in that false positive；But two
Zhou Hou, along with disease progression, Finding case liver abscess, is diagnosed as Klebsiella Pneumoniae liver abscess, thus eliminate specificity because of
Sub-false positive results；This case also illustrates, atopen has the effect of early diagnosis Klebsiella Pneumoniae liver abscess.
The positive rate of each label and false positive rate in table 3 blood culture with positive bacteria group
PagO PCR positive rate in the hemoculture sample of Klebsiella Pneumoniae liver abscess patient is 97.37%, in non-pneumonia
Positive rate in klebsiella liver abscess patient's hemoculture sample is only 0%, and the positive rate in hemoculture feminine gender group is 0%；
LuxR PCR positive rate in the hemoculture sample of Klebsiella Pneumoniae liver abscess patient is 92.11%, at non-Klebsiella Pneumoniae
Positive rate in liver abscess patient's hemoculture sample is 0%, and the positive rate in hemoculture feminine gender group is 0%；PagO+luxR is at lung
In the hemoculture sample of scorching klebsiella liver abscess patient, PCR positive rate is 97.37%, in non-Klebsiella Pneumoniae liver abscess
Positive rate in patient's hemoculture sample is only 0%, and the positive rate in hemoculture feminine gender group is 0%.These results suggest that pagO
In Klebsiella Pneumoniae liver abscess patient's blood culture with positive bacteria specimen, there is the highest specificity with luxR, can distinguish or combine
Molecular marker as Klebsiella Pneumoniae liver abscess quick diagnosis.
SEQUENCE LISTING
<110>Nanjing No.1 Hospital
<120>a kind of cause liver abscess Klebsiella Pneumoniae molecular detection kit and application thereof
<130>a kind of cause liver abscess Klebsiella Pneumoniae molecular detection kit and application thereof
<160> 6
<170> PatentIn version 3.1
<210> 1
<211> 903
<212> DNA
<213>artificial sequence
<400> 1
atgcgtagaa ttactatatt tatattattc ctgttagttg ctgtaacttg gggaaccacg 60
tggttagcta tgaagattgc tcttgaaact atccctccag tctttgctac cgggatgaga 120
tttttatttt ccgctccatt attaatcatt atcgcatggg taaaaaaaat accaatttta 180
tttcctgttg gccagcgtct atttcaactt gcaattagta tgttttattt tgccattcct 240
ttctctctaa tgatttatgg tgaggtttat gtaaacccag gacttgccgc tattatattt 300
gcaaatatgc cggtggctat tttaatagca tcatttttat ttttgaatga aaaaacaaac 360
tcaatacaga ttactggact aattatcgca ttagcttcac tttcgttcat cctcatcacg 420
gaggcgcggg aaaggacaga aagccagtgg acgggagtta ttgccctttc ttctgcagta 480
ataattcatg ccatagtata tactcaatgt aaaaaaagat gctgtaaagt ctctgttata 540
tcgtttaatg cgttaccgtg ttttatagct ggggttattc tttcgctggt ggggagtatc 600
tttgagaggc cacaattatc agctttatct ttacactcta cattagcaac aatgtattta 660
ggctgttttg ctggagtttt tgggatactg tgctattttt ctctgcagaa aagggctagc 720
gctttccagg cttcgctcgt atttctcatc ttccctctaa ttgcagtaag cttggaaagt 780
tatatatatg ggaaaaccat atcaacatac tcaatcttac tgattatccc cctcgtcatt 840
ggaatactta tcacacttat ccccaaaaaa actgtcgttg ataaaaacaa aatgaatagc 900
tga 903
<210> 2
<211> 552
<212> DNA
<213>artificial sequence
<400> 2
atgaaattgt gtgtcgttac aaataataat tatttctttg ccggcatgga acatattttc 60
tcagaggttc aatgctgtct ttgtagaata tcgagttatg atgtatatgc atgcactcct 120
aattcaaacg taatcatttt attggatggt gtgaatcaca aagtttcgat aaaagagtat 180
agctatctaa aaaaaatagg tttacctgtt ttttttattc tgaacacaaa ctgtaatgtt 240
aattccaccc tcatcggaat taacatcatt aacgcacgcg aggctatcac tattttaaaa 300
gatagaatga tttctctctt taatgggggg gggcagcttg attataaacc aataaatttg 360
acaaagaagg agtcttttat tcttagatta tatatagatg gattatcatt aacacagata 420
agtgaaaaaa cagccattag aaaaaaaaca cttataactc acacacgaaa tattcttaat 480
aagaccggtg taaaacacca taatcacctt aatattttaa aaaacatcct tggcatacat 540
ttggctcatt aa 552
<210> 3
<211> 21
<212> DNA
<213>artificial sequence
<400> 3
tgctcttgaa actatccctc c 21
<210> 4
<211> 18
<212> DNA
<213>artificial sequence
<400> 4
ggcaataact cccgtcca 18
<210> 5
<211> 20
<212> DNA
<213>artificial sequence
<400> 5
ctttgccggc atggaacata 20
<210> 6
<211> 22
<212> DNA
<213>artificial sequence
<400> 6
tgagccaaat gtatgccaag ga 22
Claims (9)
1. one kind causes liver abscess Klebsiella Pneumoniae gene specific sequencepagO, described gene specific sequencepagO
Shown in SEQ ID NO.1.
2. one kind causes liver abscess Klebsiella Pneumoniae gene specific sequenceluxR, described gene specific sequenceluxR
Shown in SEQ ID NO.2.
3. based on the primer of specific sequence design described in claim 1.
Primer the most according to claim 3, it is characterised in that:pagO_ F:TGCTCTTGAAACTATCCCTCC,pagO_ R:GGCAATAACTCCCGTCCA.
5. based on the primer of specific sequence design described in claim 2.
Primer the most according to claim 5, it is characterised in that:luxR_ F:CTTTGCCGGCATGGAACATA,luxR_ R:TGAGCCAAATGTATGCCAAGGA.
7. the specific sequence described in claim 1 or the primer described in claim 3 cause liver abscess kerekou pneumonia primary in preparation
Application in bacterium detection kit.
8. the specific sequence described in claim 2 or the primer described in claim 5 cause liver abscess kerekou pneumonia primary in preparation
Application in bacterium detection kit.
9. cause a liver abscess Klebsiella Pneumoniae molecular detection kit, including:
pagO_ F:TGCTCTTGAAACTATCCCTCC
pagO_ R:GGCAATAACTCCCGTCCA
luxR_ F:CTTTGCCGGCATGGAACATA
luxR_ R:TGAGCCAAATGTATGCCAAGGA.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610669762.XA CN106011154B (en) | 2016-08-15 | 2016-08-15 | A kind of cause hepatapostema Klebsiella Pneumoniae molecular detection kit and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610669762.XA CN106011154B (en) | 2016-08-15 | 2016-08-15 | A kind of cause hepatapostema Klebsiella Pneumoniae molecular detection kit and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106011154A true CN106011154A (en) | 2016-10-12 |
| CN106011154B CN106011154B (en) | 2019-05-31 |
Family
ID=57134844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610669762.XA Active CN106011154B (en) | 2016-08-15 | 2016-08-15 | A kind of cause hepatapostema Klebsiella Pneumoniae molecular detection kit and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106011154B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058511A (en) * | 2017-03-01 | 2017-08-18 | 深圳市宝安区沙井人民医院 | One kind sets up the triple qPCR methods of Klebsiella Pneumoniae in quick detection respiratory tract sample |
| CN110669852A (en) * | 2019-11-04 | 2020-01-10 | 深圳市人民医院 | Kit for detecting high-toxicity non-mucus Klebsiella pneumoniae |
| CN116808163A (en) * | 2023-06-16 | 2023-09-29 | 南京市中医院 | Traditional Chinese medicine composition for clearing lung and toxin, preparation method thereof and traditional Chinese medicine and western medicine composition containing same |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110287452A1 (en) * | 2010-05-18 | 2011-11-24 | National Defense Medical Center | Test Kit for Detecting Klebsiella Pneumoniae Serotype K1 and Method Using Same |
| CN103160574A (en) * | 2011-12-09 | 2013-06-19 | 中山大学达安基因股份有限公司 | Klebsiella pneumoniae nucleic acid detection kit (PCR-fluorescent probe method) |
-
2016
- 2016-08-15 CN CN201610669762.XA patent/CN106011154B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110287452A1 (en) * | 2010-05-18 | 2011-11-24 | National Defense Medical Center | Test Kit for Detecting Klebsiella Pneumoniae Serotype K1 and Method Using Same |
| CN103160574A (en) * | 2011-12-09 | 2013-06-19 | 中山大学达安基因股份有限公司 | Klebsiella pneumoniae nucleic acid detection kit (PCR-fluorescent probe method) |
Non-Patent Citations (1)
| Title |
|---|
| CHI-TAI FANG等: "A Novel Virulence Gene in Klebsiella pneumoniae Strains Causing Primary Liver Abscess and Septic Metastatic Complications", 《THE JOURNAL OF EXPERIMENTAL MEDICINE》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058511A (en) * | 2017-03-01 | 2017-08-18 | 深圳市宝安区沙井人民医院 | One kind sets up the triple qPCR methods of Klebsiella Pneumoniae in quick detection respiratory tract sample |
| CN110669852A (en) * | 2019-11-04 | 2020-01-10 | 深圳市人民医院 | Kit for detecting high-toxicity non-mucus Klebsiella pneumoniae |
| CN116808163A (en) * | 2023-06-16 | 2023-09-29 | 南京市中医院 | Traditional Chinese medicine composition for clearing lung and toxin, preparation method thereof and traditional Chinese medicine and western medicine composition containing same |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106011154B (en) | 2019-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ikryannikova et al. | Misidentification of alpha-hemolytic streptococci by routine tests in clinical practice | |
| CN104946762B (en) | Detect the kit of Friedlander's bacillus | |
| CN103361434A (en) | Multiple fluorescence PCR detection kit and detection method for clostridium difficile toxin genes | |
| Zhou et al. | Using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) complemented with selected 16S rRNA and gyrB genes sequencing to practically identify clinical important viridans group streptococci (VGS) | |
| Abraham et al. | Identification of Streptococcus pyogenes–Phenotypic Tests vs Molecular Assay (spy1258PCR): A Comparative Study | |
| Li et al. | Modulation of gut microbiota, short-chain fatty acid production, and inflammatory cytokine expression in the cecum of porcine deltacoronavirus-infected chicks | |
| CN106011154B (en) | A kind of cause hepatapostema Klebsiella Pneumoniae molecular detection kit and its application | |
| CN105779625A (en) | Dual-fluorescence quantitative PCR (Polymerase Chain Reaction) primer, kit and method for simultaneously detecting general type and type 2 Streptococcus suis | |
| CN105525023A (en) | Fluorescent quantitative PCR detection kit for clostridium difficile toxin A/B and detection method | |
| CN102776283A (en) | Visualized loop-mediated isothermal amplification kit for detecting Haemophilus parasuis | |
| CN107142331A (en) | A kind of primer and probe for being used to detect mycoplasma ovine pneumoniae | |
| CN102242216A (en) | Fluorescent PCR (polymerase chain reaction) detection kit for vibrio cholerae and systematic identification method thereof | |
| Ohud et al. | Detection of Salmonella strains in clinical samples from Saudi Arabia by invA and hilA polymerase chain reaction (PCR)-based assays | |
| CN104561277B (en) | For detecting the target sequence and detection kit of mycoplasma pneumoniae | |
| CN106801103B (en) | Detection primer group, detection kit and multiplex PCR detection method for streptococcus agalactiae | |
| CN111118182B (en) | Specific novel molecular target of serotype listeria monocytogenes and rapid detection method thereof | |
| Zaheer et al. | High-throughput species identification of enterococci using pyrosequencing | |
| Yang et al. | Rapid identification of bacterial species associated with bronchiectasis via metagenomic approach | |
| CN103882140A (en) | Rapid diagnosis kit for mycobacterium tuberculosis | |
| Wang et al. | Multilocus sequence typing and virulence analysis of Haemophilus parasuis strains isolated in five provinces of China | |
| CN106544432A (en) | A kind of drug resistance of Staphylococcus aureus and virulence method for quick and test kit | |
| CN103305613B (en) | Giant salamander pathogenic hydrophila gingivalis PCR diagnostic kit | |
| CN107988400A (en) | Detect the reagent set of staphylococcus haemolyticus | |
| CN104846067A (en) | Duplex PCR detection kit and detection method for Listeria monocytogenes and Enterococcus faecium | |
| CN103122383B (en) | Horizontal fluorescent quantitation polymerase chain reaction (PCR) detection method of streptococcus suis strain |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |