E coli fluorescence kuaj PCR lub sijhawm tiag tiag

Lub sijhawm tiag tiag fluorescent PCR hauv vitro kuaj pom ntawm E coli.

Kev qhia tshwj xeeb: 96 T / lub thawv
Cov khoom siv tau khaws cia ntawm 2 - 8 ℃.
Lub sijhawm siv tau yog 12 lub hlis.

specification: 96T / lub thawv

Siv: Lub sijhawm tiag tiag fluorescent PCR hauv vitro kuaj pom ntawm e coli.

Txoj cai tshawb nrhiav: Cov khoom siv muaj cov kab mob tshwj xeeb e coli. Thaum cov qauv muaj e coli, e coli DNA muab rho tawm tom qab kev ua kom zoo ntxiv tau nthuav dav los ntawm PCR. Thaum lub sijhawm tshuaj tiv thaiv, e-coli-tshwj xeeb fluorescent sojntsuam sib xyaw nrog lub hom phiaj nucleic acid thiab ib txhij Taq enzyme Hydrolysis txhawm rau cais cov pab pawg fluorescent los ntawm pab pawg quenching ntawm kev sojntsuam: yog li muaj cov e coli tuaj yeem raug txiav txim hauv lub sijhawm tiag tiag qhov nce ntawm fluorescence.

Khoom thiab muaj pes tsawg leeg:

Muaj pes tsawg leeg (96T/khoom siv)	specification	kom muaj nuj nqis
Kev daws cov tshuaj DNA	3ml	1 raj
E coli PCR cov tshuaj tiv thaiv tshuaj	1100ul	1 raj
Kais enzyme (5U/ul)	20ul	1 raj
E coli zoo tswj	40ul	1 raj
E coli Tsis Zoo Tswj	0.5ml	1 raj
phau ntawv qhia		1

Cov xwm txheej cia thiab hnub tas sij hawm ntawm cov khoom siv:
1. Cov khoom siv tau khaws cia ntawm 2 - 8 ℃.
2. Lub sijhawm siv tau yog 12 lub hlis.

Cov cuab yeej siv tau:
ABI7500, 9600 series, MJ opticon2, Bio-Rad thiab lwm yam fluorescent ntau qhov ntsuas PCR muaj

Txoj kev sim:
1. Siv 1ml ntawm cov tshuaj ua kom zoo rau hauv 1.5m1 cov menyuam tsis muaj menyuam centrifuge raj, centrifuge ntawm 10.000rpm rau 5min, thiab pov tseg cov supernatant; siv cov txheej txheem rho tawm phenol chloroform qub los kuaj cov ntawv rho tawm lossis khaws nws ntawm 20C kom txog thaum nws raug sim.
2. Muab PCR cov tshuaj tiv thaiv ntawm chav sov kom sib npaug, coj Taq enzyme tom qab yaj, thiab ntxiv Taq enzyme ntawm 2∪/raj, uas yog, txhua qhov kev sim tshuaj tiv thaiv kab mob tau npaj ua: PCR cov tshuaj tiv thaiv tshuaj 22.5u1+0.4ulTaq enzyme.
3. Ntxiv tus qauv: Thaw cov nucleic acid extract khaws cia ntawm -20 ° C ntawm chav sov thiab centrifuge ntawm 13,000 rpm rau 5 feeb. Qhov tsis zoo thiab kev tswj hwm tau yaj hauv chav sov ua ntej siv. Hauv txhua lub PCR cov tshuaj tiv thaiv raj, ntxiv 2 ul ntawm tus qauv lossis qhov tsis zoo thiab kev tswj hwm tau ua tiav hauv qib 1, npog lub raj, tso rau ntawm PCR ntsuas, thiab sau tus lej piv txwv.
4. On-board amplification thiab nrhiav chaw: qhov xwm txheej tshwm sim yog 95C: 3min, 1 lub voj voog; 95C: 5sec, 60C: 40sec (teeb liab tau txais), 40 mus. Cov tshuaj tiv thaiv kab mob tau teeb tsa rau 25u1.
Rau ntau-channel fluorescence PCR ntsuas, lub teeb liab tau teeb tsa rau Fam fluorescein.

Cov txiaj ntsig kev txiav txim:
Thov mus saib phau ntawv qhia kom ntxaws.

Ceev faj rau kev siv cov khoom siv:
1. Cov khoom siv no tsuas yog siv rau hauv vitro kuaj thiab tshawb fawb lub hom phiaj, thov nyeem tag nrho cov ntawv ntawm phau ntawv no ua tib zoo ua ntej pib siv nws.
2. Kev sim yuav tsum tau muab faib nruj me ntsis: thaj tsam npaj ua ntej PCR-npaj cov tshuaj reagents; cheeb tsam ua qauv-yuav tsum kuaj cov qauv, ua qauv; thaj tsam nrhiav pom-PCR kev nthuav dav, nrhiav pom.
3. Rau cov txheej txheem kev tswj hwm hauv chaw sim, thov saib mus rau qhov ua tiav ntawm cov qauv kev tswj hwm rau kev hloov pauv cov noob tso tawm los ntawm cov koom haum hauv tebchaws.

Yog tias koj xav paub ntau ntxiv, thov hu rau peb

Cov lus nug tsis tseem ceeb: E coli

Txog e coli

Ua coli, tseem hu ua Escherichia coli, tau tshawb pom los ntawm Escherich xyoo 1885. Escherichia coli yog ib hom kab mob uas muaj kab mob. Hauv qee qhov xwm txheej, nws tuaj yeem ua rau mob plab zom mov lossis kab mob tso zis ntawm ntau cov ntaub so ntswg thiab kabmob hauv tib neeg thiab ntau yam tsiaj.

E coli loj

Qhov loj yog 0.5 × 1 ～ 3 microns. Chow flagella, tuaj yeem txav mus yam tsis muaj kab mob. Nws tuaj yeem ferment ntau yam suab thaj los ua kua qaub thiab roj. Nws yog cov kab mob ib txwm nyob hauv cov hnyuv ntawm tib neeg thiab tsiaj. Nws nkag mus rau hauv cov hnyuv nrog cov menyuam mos tom qab yug los thiab nrog tib neeg nyob rau lub neej, suav txog yuav luag 1/3 ntawm qhov hnyav qhuav ntawm cov quav.

E coli puab

E coli yog Brevibacterium, nrog rau qhov xaus puag ncig thiab Gram tsis zoo. Qee zaum, vim muaj ib puag ncig sib txawv, cov tib neeg lub cev zoo li tus kheej lossis zoo ib yam li cov ntsiab lus; E. coli feem ntau yog ib leeg lossis ob, tab sis lawv yuav tsis raug teeb tsa hauv cov saw ntev; feem ntau E. coli hom muaj tsiav tshuaj lossis microcapsules Cov Qauv, tab sis tsis tuaj yeem tsim cov kab mob; feem ntau cov kab mob E. coli loj hlob tuaj fimbriae, qee qhov yog tus tswv tsev tshwj xeeb fimbriae uas muaj cov nplaum rau tus tswv tsev thiab qee cov ntaub so ntswg lossis hlwb.

E coli kev faib tawm

Cov kab mob pathogenic
Tam sim no, kev lees paub thoob ntiaj teb feem ntau suav nrog rau hom E. coli, uas yog cov kab mob hauv lub cev E. coli (EPEC), hnyuv hnyuv toxigenic E. coli (ETEC), thiab kev cuam tshuam hauv plab uas tuaj yeem ua rau mob plab zom mov. E coli (EIEC), plab hnyuv hemorrhagic E coli (EHEC), plab hnyuv sib koom ua ke E coli (EAEC) thiab hnyuv Shiga-zoo li cov co toxins pom nyob rau xyoo tsis ntev los no kuj muaj qee tus kab mob E coli (ESIES) Ntxiv rau, muaj cov kab mob urethral E coli ( UPEC) uas tuaj yeem ua rau cov kab mob tso zis, thiab lub npe tshiab hu ua hnyuv hnyuv sib koom ua ke E. coli (EAggEC).

Hemolytic
Raws li lub peev xwm ntawm E. coli los tsim hemolysin thiab muaj peev xwm hemolyze, E. coli tuaj yeem faib ua ob pawg: hemolytic E. coli thiab tsis-hemolytic E. coli.

Enterotoxogenic
Raws li lub peev xwm ntawm E coli los tsim enterotoxin thaum kis tus kab mob, E coli tuaj yeem faib ua ob pawg: enterotoxigenic E coli thiab non-entotoxinogenic E. coli. Enterotoxigenic E. coli yog kab mob tseem ceeb ntawm kev kis mob raws plab hauv tib neeg thiab ntau yam tsiaj. Kev txheeb xyuas tus kab mob enterotoxigenic E. coli feem ntau yog txhawm rau txiav txim siab hom enterotoxin zais los ntawm kev cais E. coli. Ib qho ntxiv, raws li lub peev xwm ntawm E. coli los tsim enterotoxin, ua ke nrog nws qhov kev nkag siab sib txawv enterotoxins, E coli tuaj yeem cais ua hom enterotoxin.

Tus kab mob E coli

E. coli yog cov kab mob ib txwm nyob hauv cov hnyuv ntawm cov tsiaj, thiab ib feem me ntawm lawv ua rau muaj kab mob nyob rau qee yam mob. Serotypes ntawm E. coli tuaj yeem ua rau mob plab zom mov hauv tib neeg lossis tsiaj, feem ntau yog tshwm sim los ntawm fimbria antigens tshwj xeeb, cov kab mob hauv lub cev thiab lwm yam kab mob. Ntxiv nrog rau kev mob plab zom mov, lawv kuj tuaj yeem ua rau cov kab mob tso zis, mob caj dab, thiab mob qog noj ntshav. Kev mob thiab mob septic.
Escherichia coli suav nrog ntau pawg kab mob uas muaj cov kab mob sib txawv. Tib neeg kab mob E. coli tau muab faib ua ob hom: kab mob hauv plab thiab kis kab mob sab hauv.

Cov tsos mob E coli

Cov tsos mob feem ntau ntawm tus kab mob hauv plab yuav yog raws plab. Yog tias kis tau tus mob hnyav, yuav muaj kev poob siab, ntshav siab yuav poob, thiab lub plawv dhia tuaj yeem nce ntxiv. Yog tias nws yog kab mob hauv plab, xeev siab, ntuav, ua npaws, tso zis ntau zaus, thiab xwm txheej ceev tuaj yeem tshwm sim tom qab kis kab mob E. coli.

E coli kab mob

Feem ntau, E. coli tau muab faib ua plaub pawg kab mob raws li nws cov kab mob rau tib neeg: hom kab mob E. coli, enteropathogenic E. coli, enterotoxigenic E. coli, thiab tus kab mob E. coli. Peb tom kawg yog cov kab mob pathogenic uas ua rau raws plab.

E coli pathogenesis

Txhua lub paj suav nrog qee tus naj npawb ntawm serotypes. Enterotoxigenic Escherichia coli tuaj yeem tsim cov cua kub-resistant enterotoxin thiab cua sov-resistant enterotoxin. Kev tsim tawm ntawm ob lub enterotoxins no yog kev tswj hwm los ntawm plasmids. Plasmid yog ib hom DNA tsis-chromosomal uas tuaj yeem kis tau yooj yim. Kev sib kis sib kis ntawm plasmids tuaj yeem ua rau cov kab mob tsis-lom ua kom tau txais cov peev txheej tsim cov tshuaj lom. Vim yog kev xa cov plasmid, Escherichia coli ntawm ib qho serotype tuaj yeem tsim cov enterotoxin thiab cov neeg nyob hauv pawg, yog li muaj cov kab mob.

Kev kho mob E coli

Ua ntej, yog tias cov tsos mob ntawm kev kis mob tsis hnyav heev thiab raws plab thiab mob plab me me, kev tswj qhov ncauj ntawm bifidobacteria tuaj yeem txhim kho txoj hnyuv zom zaub mov tsis zoo thiab tswj cov plab hnyuv microflora.
Yog tias mob plab thiab raws plab hnyav thiab muaj quav dej, txiav txim siab muab tshuaj tiv thaiv kab mob xws li tshuaj tua kab mob quinolone lossis berberine, thiab hloov dej kom tsim nyog los txhim kho cov tsos mob lub cev qhuav dej.

E coli tuaj qhov twg tuaj

Escherichia coli yog ib qho ntawm cov kab mob me me tshaj plaws, tau nthuav dav nyob rau hauv qhov, feem ntau tsis yog kab mob, feem ntau yog kab mob hauv cov hnyuv lossis tib neeg lossis tsiaj, thiab hu ua "ib txwm muaj paj" los ntawm tib neeg.

E coli kis tau tus mob

Lub hauv paus tseem ceeb ntawm kev kis mob rau tib neeg tus kab mob E. coli yog tias muaj coob tus kab mob E. coli raug tso tawm hauv cov quav ntawm cov neeg mob plab zom mov. Txoj kev sib kis ntawm tus kab mob E. coli ntawm tib neeg feem ntau yog los ntawm txoj kev kis kab mob hauv lub qhov ncauj, uas tuaj yeem ua rau kis lossis kis tus kab mob E. coli nyob rau qee qhov xwm txheej.

Enterobacter sakazakii hauv cov mis mos ua hmoov

Enterobacter sakazakii yog tsiaj txhu uas muaj nyob hauv ib puag ncig thiab feem ntau pom muaj nyob hauv cov tsev thiab ib puag ncig hauv tsev uas cov mis mos tau tsim. Txawm hais tias tus menyuam mos mis tsis yog cov khoom lag luam uas tsis muaj menyuam thiab tej zaum yuav kis tau cov kab mob uas tuaj yeem ua rau muaj kab mob loj, yog tias tus menyuam mos mis tau npaj thiab khaws cia kom raug, kev pheej hmoo kis mob los ntawm Enterobacter sakazakii tuaj yeem txo qis. Txhawm rau npaj cov mis mos kom nyab xeeb, koj yuav tsum xub ua kom huv thiab tsis muaj menyuam kom pub mis thiab npaj khoom siv.

Kev tiv thaiv kab mob E coli

1. Khaws cov bai qhov chaw thiab tais diav hauv chav ua noj kom huv, thiab muab pov tseg kom huv.
2. Khaws koj txhais tes kom huv thiab txiav koj cov rau tes ntau zaus.
3. Ntxuav tes nrog xab npum thiab dej ua ntej noj mov lossis tuav zaub mov. Koj yuav tsum tau ntxuav koj txhais tes tom qab siv chav dej lossis hloov ntaub pawm.
4. Cov kais dej yuav tsum tau siv rau cov dej haus, thiab nws yog qhov zoo tshaj los rhaub nws ua ntej haus.
5. Cov zaub mov tshiab yuav tsum tau yuav los ntawm qhov chaw ntseeg tau, tsis yog tus saib xyuas tsis muaj ntawv tso cai.
6. Tsis txhob noj zaub mov uas muaj kev pheej hmoo siab, xws li mis nyuj uas tsis tau ua tiav los ntawm qhov ntsuas kub qis kom tsis muaj menyuam, nrog rau cov nqaij hamburger tsis qab, cov nqaij hauv av, thiab lwm yam zaub mov.
7. Thaum ua zaub mov noj, hnav kom huv, phuam qhwv thiab kaus mom.
8. Cov zaub mov yuav tsum tau ntxuav kom huv.
9. Cov khoom noj tsis zoo yuav tsum tau npog nrog lub hau thiab muab cia rau hauv lub tub yees.
10. Cov zaub nyoos thiab cov zaub mov siav, tshwj xeeb yog nqaij nyuj thiab nqaij nyuj offal, yuav tsum tau tuav thiab khaws cia cais (khaws cov zaub mov siav rau saum lub tub yees thiab cov zaub nyoos rau hauv qab) kom tsis txhob kis kab mob sib kis.
11. Lub tub yees yuav tsum tau ntxuav thiab yaj tsis tu ncua, thiab qhov kub yuav tsum tau khaws cia ntawm 4 degrees Celsius lossis qis dua.
12. Yog tias txhua feem ntawm cov zaub mov raug kub txog 75 degrees Celsius, E. coli O157: H7 tuaj yeem tshem tawm; yog li ntawd, nqaij nyuj hauv av thiab hamburger steak yuav tsum tau ua kom huv si kom txog 75 degrees Celsius rau 2 mus rau 3 feeb, kom txog thaum cov nqaij siav tag nrho tig mus Nws yog xim av, thiab cov gravy ua kom pom tseeb.
13. Tsis txhob tuav cov zaub mov siav nrog txhais tes liab qab; yog tias tsim nyog, looj hnab looj tes.
14. Cov zaub mov yuav tsum tau noj sai li sai tau tom qab ua noj.
15. Yog tias nws tsim nyog khaws cov zaub mov uas seem, nws yuav tsum tau muab tso rau hauv tub yees thiab noj kom sai li sai tau. Reheat kom huv ua ntej noj mov. Cov zaub mov lwj yuav tsum muab pov tseg.

Test rau e coli

Txoj kev fermentation

Hom no feem ntau yog rau kev cog qoob loo ntawm E. coli ntawm qhov nruab nrab ntawm 44.5 ° C, uas muaj cov tshuaj fluorescent thiab xav tau 24 teev ntawm kev cog qoob loo. Cov kauj ruam tseem ceeb suav nrog fermentation, cais kev coj noj coj ua, theem nrab fermentation, tsom tsom tsom iav, thiab lwm yam.

Matrix-pab laser desorption ionization lub sij hawm-ntawm-dav hlau huab hwm coj spectrometry

Matrix-pab laser desorption ionization time-of-flight mass spectrometry (MAL-DITOF-MS) txoj kev yog sib xyaw cov qauv coj los ntsuas nrog cov tshuaj matrix ntawm qee qhov sib piv, thiab siv cov tshuab hluav taws xob pulsed kom ua rau lub matrix, ua rau polymer los tsim cov tshuaj molecular ions thiab molecular ion polymerization Lub cev tau nrawm dua thiab kuaj pom nyob rau lub sijhawm-ntawm-kev ya dav hlau loj spectrometer. Raws li qhov tshwm sim, ntsuas polymer cais raws li qhov huab hwm coj tus lej, ua tiav los tsim cov pawg loj, thiab tshuaj xyuas thiab sib piv los ntawm cov software los tshuaj xyuas thiab txiav txim siab qhov tshwj xeeb (ntiv tes) spectrum kom ua tiav qhov sib txawv thiab txheeb xyuas cov hom phiaj kab mob me me lossis kab mob.

Immunomagnetic hlaws txoj kev

Lub hauv paus ntsiab lus ntawm kev sib cais thev naus laus zis yog siv cov hlaws sib nqus ua tus nqa khoom thiab cov tshuaj tiv thaiv los ua ke cov tshuaj tiv thaiv kab mob thiab cov hlaws sib nqus, thiab tom qab ntawd ua kom tiav cov tshuab txav los ntawm kev siv tshuab sib nqus kom cais E. coli. Piv nrog rau lwm txoj hauv kev cais cov kab mob, txoj hauv kev no muaj qee qhov zoo. Cov thev naus laus zis no tuaj yeem txhim kho kev txheeb xyuas qhov ua tiav ntawm cov kab mob Vibrio hauv cov qauv, thiab cov thev naus laus zis thev naus laus zis tuaj yeem kho cov kab mob sib txawv hauv cov kab mob sib txawv. , Thiab mus rau qhov loj txhim kho qhov ua kom pom tseeb.

Kev ntsuas ntsuas qhov ntsuas

Qhov tseem ceeb ntawm kev siv tshuaj tiv thaiv kab mob tiv thaiv kab mob, kev siv thev naus laus zis no tau tsim thiab siv hauv xyoo 1970. Nrog kev txhim kho thiab kev nce qib ntawm kev tshawb fawb thiab thev naus laus zis, kev siv lub tshuab ntsuas ntsuas tsis siv neeg tau siv dav, thiab nws yooj yim heev rau kev khiav lag luam, nws tuaj yeem txuag sijhawm ntau. , nws qib kev cuam tshuam yog me me, nws tuaj yeem txuag cov neeg ua haujlwm thiab cov khoom siv nkag, thiab tseem tuaj yeem txhim kho qhov tseeb ntawm kev tshawb nrhiav. Hauv cov txheej txheem kev txhim kho tam sim no, kev thov tsis siv neeg enzyme immunoassay system yog qhov dav heev.

ATP bioluminescence

Hauv txoj kev txhim kho hauv xyoo tas los no, thev naus laus zis bioluminescence tau siv dav, thiab nws yog thev naus laus zis nrawm rau tshuaj xyuas kab mob. Hauv cov hlwb nquag, ATP yog nws cov khoom siv hluav taws xob siv hluav taws xob ntau, uas tuaj yeem muab lub zog xav tau thaum lub sijhawm ua haujlwm ntawm lub cev. Thiab, cov thev naus laus zis no tuaj yeem tswj hwm qee cov ntsiab lus hauv qee yam hauv lub cev. Kev tshawb pom thev naus laus zis ntawm Escherichia coli hauv cov zaub mov tuaj yeem siv txoj hauv kev ntawm cov duab fluorescence, vim tias vim li cas lub cev tso tawm lub teeb yog lub luag haujlwm ntawm luciferase, uas ua rau muaj txiaj ntsig ntawm lub teeb tawm. Cov tshuaj tau los ntawm cov foob pob hluav taws hauv North America thiab tuaj yeem cuam tshuam qhov oxidation ntawm fluorescein. Txawm li cas los xij, cov tshuaj tsis ruaj khov thiab tuaj yeem rhuav tshem cov tshuaj fluorescence sai. Ib qho ntxiv, kev txheeb xyuas thev naus laus zis cov txiaj ntsig tau txais cov txheej txheem tau nrawm heev, thiab cov cuab yeej nqa tau yooj yim, tsim nyog rau ntawm qhov chaw tshawb nrhiav.