Salmonella Fluorescence real-time PCR test

Real-time fluorescent PCR hauv vitro nrhiav pom ntawm Salmonella.

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: Real-time fluorescent PCR hauv vitro nrhiav pom ntawm Salmonella.

Txoj cai tshawb nrhiav: Cov khoom siv muaj Salmonella tshwj xeeb primers. Thaum cov qauv muaj Salmonella, Salmonella DNA tau muab rho tawm tom qab kev txhim kho ua ntej yog exponentially amplified los ntawm PCR. Thaum cov tshuaj tiv thaiv, Salmonella tshwj xeeb fluorescent sojntsuam hybridizes nrog lub hom phiaj nucleic acid thiab ib txhij Taq enzyme Hydrolysis cais pawg fluorescent los ntawm pawg quenching ntawm qhov kev sojntsuam: kom lub xub ntiag ntawm Salmonella tuaj yeem txiav txim siab ntawm lub sijhawm los ntawm fluorescence. nce.

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
Salmonella PCR tshuaj tiv thaiv	1100ul	1 raj
Kais enzyme (5U/ul)	20ul	1 raj
Salmonella zoo tswj	40ul	1 raj
Salmonella Negative Control	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 cov qauv: Thaw cov nucleic acid extract khaws cia ntawm -20 ° C ntawm chav tsev kub thiab centrifuge ntawm 13,000 rpm rau 5 feeb. Cov kev tswj tsis zoo thiab qhov zoo yog thawed ntawm chav tsev kub ua ntej siv. Hauv txhua lub PCR cov tshuaj tiv thaiv raj, ntxiv 2 ul ntawm cov qauv lossis cov kev tswj tsis zoo thiab cov kev tswj xyuas zoo hauv kauj ruam 1, npog lub raj, tso rau ntawm PCR ntsuas, thiab sau tus lej coj mus kuaj.
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: Salmonella

Hais txog Salmonella

Salmonella txhais

Salmonella yog ib lo lus dav dav rau ib pawg kab mob. Nws tuaj yeem kis tau rau tib neeg thiab ua rau muaj kab mob thiab tsiaj txhu. Nws yog tus kab mob zoonotic. Vim Salmonella yog Gram-negative Enterobacteriaceae, tib neeg tuaj yeem noj zaub mov uas muaj kab mob Salmonella thiab ua rau cov zaub mov lom. Raws li kev txheeb cais, nyob rau hauv lub ntiaj teb no ntawm cov zaub mov lom, feem ntau yog tshwm sim los ntawm Salmonella noj zaub mov yog ib tug ntau yam khoom noj kab mob. Tsis tas li ntawd, Salmonella typhimurium, Salmonella paratyphi, Salmonella choleraesuis, thiab lwm yam, tuaj yeem ua rau mob plab hnyuv xws li mob plab, mob plab paratyphoid, thiab kab mob cholera. Tus kab mob typhoid tsuas kis tau rau tib neeg, tsis yog tsiaj txhu, thiab ua rau tib neeg cov pob txha pob txha, daim siab thiab tus po. Txhua xyoo, nws tuaj yeem ua rau kaum lab tus tib neeg mob thiab ntau dua 600,000 tuag.

Salmonella qhov chaw

Salmonella muaj ntau yam ntawm cov tswv hauv qhov xwm txheej. Ob peb Salmonella yog xaiv rau tus tswv tsev, thiab lawv feem ntau haum rau tib neeg thiab tsiaj txhu. Lawv tuaj yeem nyob hauv lub plab zom mov ntawm cov tsiaj txhu, tsiaj reptiles, noog, kab, thiab tib neeg. Tus kab mob kis nrog cov tsiaj qus yog ntau tshaj 1% txog 20%.

Salmonella feem ntau ua rau cov nqaij noj nqaij, thiab ntses, nqaij qaib, mis nyuj, thiab qe zaub mov kuj tuaj yeem kis tau los ntawm cov kab mob no. Salmonella zaub mov lom tuaj yeem tshwm sim thoob plaws hauv lub xyoo. Noj cov kab mob uas tsis tau noj, cov nqaij tsiaj txhu tuag, lossis cov nqaij tsiaj txhu uas muaj kab mob ntawm lwm cov ntsiab lus tom qab tua yog qhov tseem ceeb tshaj plaws ua rau muaj kab mob salmonella. Qee cov kws paub txog zaub mov taw qhia tias cov neeg Asmeskas muaj kev noj zaub mov txawv ntawm cov neeg Suav. Lawv nyiam noj ib nrab-boiled qe los yog cov qe nyoos, yog li thaum lub qe muaj salmonella, txoj kev pheej hmoo ntawm kev kis kab mob kuj muaj ntau. "Hauv lub tebchaws, nws tsis yog qhov xav tsis thoob tias cov qe nyoos muaj cov kab mob salmonella, tab sis tib neeg nyiam ua cov qe siav, uas txo qis kev kis kab mob.

Kev kis tus kab mob Salmonella

Salmonella belongs rau tsev neeg Enterobacteriaceae thiab tuaj yeem ua rau gastroenteritis, typhoid fever, sepsis, thiab kab mob hauv plab focal. Nws raug hu ua tus kab mob Salmonella (xws li, salmonellosis). Ntau yam syndromes tuaj yeem sib tshooj. Cov kab mob uas tshwm sim los ntawm cov kab mob typhoid thiab paratyphoid tsis suav nrog.

Cov tsos mob ntawm tus kab mob Salmonella muaj ntau haiv neeg. Raws li lawv cov tsos mob tseem ceeb, lawv tuaj yeem muab faib ua plaub hom: enteritis, typhoid, septicemia, thiab kab mob hauv zos suppurative.

Cov kab mob Salmonella

Salmonella tus kab mob

Salmonella typhi yog cov kab mob uas muaj qhov loj ntawm (0.6 txog 0.9) × (1 txog 3) microns tsis muaj spores thiab feem ntau tsis muaj tshuaj ntsiav. Tsuas yog Salmonella gallinarum thiab Salmonella typhi, feem ntau ntawm lawv muaj flagella nyob ib ncig ntawm lub cev, ua rau Salmonella typhi.
Cov kab mob Salmonella uas ua rau mob typhoid fever. Cov kab mob zoo li tus pas nrig, nrog flagella thoob plaws lub cev thiab tuaj yeem tsim cov tshuaj endotoxin. Kev txhaj tshuaj Endotoxin rau hauv cov leeg luav tuaj yeem ua rau tib neeg zoo li cov kab mob, xws li ua npaws, leukopenia, thiab lwm yam, thiab kev txhaj tshuaj loj tuaj yeem ua rau tuag taus.

Salmonella typhimurium

Salmonella typhimurium yog ib pawg ntawm cov tsis hloov pauv lossis pantrophic salmonella. Nws muaj ntau hom tswv thiab faib rau hauv qhov xwm txheej. Nws muaj nyob rau hauv txoj hnyuv ntawm ntau yam tsiaj xws li nqaij qaib, tsiaj txhu, thiab nas. Ib. Cov kab mob tuaj yeem ua rau muaj ntau yam kab mob sib kis ntawm cov nqaij qaib thiab tsiaj txhu, thiab tuaj yeem ua rau tib neeg muaj kab mob, thiab muaj qhov tseem ceeb ntawm kev noj qab haus huv rau pej xeem.

Salmonella enteritis

Salmonella enteritidis yog cov kab mob tseem ceeb ua rau mob plab hnyuv. Cov tsos mob tshwm sim tom qab kis tau muaj xws li kub taub hau, raws plab, thiab ntuav. Salmonella enteritidis yog ib qho ntawm cov kab mob pathogenic uas tsis muaj tus tswv tsev tshwj xeeb thiab nkag mus. Tus tswv tsev suav nrog tib neeg thiab ntau yam tsiaj. Tsis tsuas yog cov kab mob tuaj yeem ua rau muaj kev puas tsuaj loj rau cov nqaij qaib thiab kev tuag ntawm cov nqaij qaib, tab sis cov khoom noj nqaij qaib uas kis tau tus kab mob Salmonella enteritidis kuj ua rau muaj kev phom sij rau tib neeg kev noj qab haus huv.

Salmonella choleraesuis

Salmonella choleraesuis yog ib hom kab mob uas tsis yog-typhoidal Salmonella, uas yog cov kab mob tseem ceeb ua rau cov kab mob paratyphoid hauv piglets, ua rau muaj kev phom sij loj rau kev lag luam npua.
Thaum tus kab mob septicaemic Salmonella choleraesuis invades lub cev ntawm tus npua, nws tam sim ntawd invades lub lymphatic chaw ntawm lub plab hnyuv phab ntsa, yog li invading cov ntshav dua thiab multiply nyob rau hauv cov ntshav, ua rau septicemia. Ntau tus endotoxins raug tsim thaum cov kab mob rov ua dua, thiab endotoxins ua rau cov qe ntshav dawb thiab ua rau mob, kub taub hau thiab raws plab.
Tus kab mob ntawm Salmonella choleraesuis feem ntau kis los ntawm faecal orifice. Yog tias tus neeg mob muaj tus npua teb hauv tsev, kev tu cev tsis zoo, thiab lub hauv paus ntawm cov kab mob siab ua haujlwm tsis zoo, kev tiv thaiv kab mob qis, yog li nws yuav yog vim kev noj zaub mov lossis dej paug rau tus kab mob Salmonella.

Tom qab tus neeg laus kis tus kab mob Salmonella choleraesuis, qhov pib yog insidious thiab kev kho mob tshwm sim tsis raug. Nws yog ib qho yooj yim kom tsis nco qab kev kuaj mob thiab misdiagnosis thiab ncua kev kho mob. Yog tias chav kawm tshuaj tsis txaus, kev tswj hwm tus kab mob insidious bacteremia tsis tiav thiab tus mob tuaj yeem ua rau ntev dua.

Salmonella lom

Tus tswv tsev tshwj xeeb ntawm Salmonella yog qhov tsis muaj zog, uas tuaj yeem kis tau ob qho tib si tsiaj thiab tib neeg, thiab nws yooj yim los ua zaub mov lom rau tib neeg. Cov kab mob feem ntau yog Salmonella cholerae, ua raws li Salmonella typhimurium thiab Salmonellaenteritidis.
Kev noj cov zaub mov uas tsis yog typhoidal Salmonella kis los ntawm qhov ncauj ua rau muaj mob. Cov khoom noj uas ua rau muaj mob feem ntau yog tsiaj txhu, feem ntau yog cov tsiaj txhu, nqaij qaib, qe, mis nyuj thiab lawv cov khoom.

Cov tsos mob Salmonella

Cov tsos mob ntawm cov zaub mov lom uas tshwm sim los ntawm salmonella feem ntau yog xeev siab, ntuav, mob plab, mob taub hau, ua daus no thiab raws plab, nrog rau qaug zog, mob nqaij, tsis pom kev, kub taub hau, tsis xis nyob thiab tsaug zog, lub sijhawm ntev yog 2 mus rau 3 hnub, qhov nruab nrab ntawm cov khoom noj muaj tshuaj lom. tuag tus nqi yog 4.1%. Qhov laj thawj tseem ceeb yog tshwm sim los ntawm kev noj cov zaub mov uas muaj ntau ntawm cov kab mob uas tsis yog tus tswv tsev lossis cov kab mob ntawm Salmonella.

Salmonella pathogenesis

1. Kev ntxeem tau ntawm cov kab mob salmonella nyob rau ntawm txoj hnyuv mucosa thiab cov nyhuv synergistic ntawm endotoxin; nws yog lub ntsiab ua rau cov zaub mov lom los ntawm salmonella.

2. enterotoxin lom: Salmonella enteritidis thiab Salmonella typhi tuaj yeem tsim enterotoxin. Lub tshuab yog tib yam li enterotoxin hom kab mob zaub mov lom.

Salmonella incubation lub sij hawm

Tom qab noj tshuaj lom. Cov tsos mob feem ntau tshwm sim li ntawm 12 mus rau 14 teev, thiab qee qhov muaj lub sijhawm incubation ntev dua.

Kev kho mob Salmonella

Gastroenteritis tshwm sim los ntawm Salmonella feem ntau tsis tas yuav siv tshuaj tua kab mob; nws yog tsuas yog supplemented nrog dej thiab electrolytes; tab sis rau neonates, cov neeg mob immunocompromised thiab tshwj xeeb raug mob; Kev kho tshuaj tua kab mob yuav tsum tau nyob ntawm qhov xwm txheej.

Kev tiv thaiv Salmonella

1. Ntxuav koj ob txhais tes kom zoo ua ntej noj mov, tom qab mus rau hauv chav dej, ua ntej kov zaub mov, tom qab kov cov tsiaj los yog qe nyoos.
2. Muab cov txiav txiav rau cov zaub mov nyoos thiab siav.
3. Noj cov zaub mov kom huv si (tshwj xeeb yog qe thiab nqaij qaib).
4. Cov zaub mov uas tsis npaj yuav noj yuav tsum muab qhwv rau hauv hnab yas thiab muab cia rau hauv lub tub yees. Nws yuav tsum tau rhaub los yog siav ua ntej noj dua.
5. Sib ntaus thiab thaiv cov vectors xws li yoov. Cov zaub mov tsis huv uas muaj kab mob yoov, tas sij hawm lossis lwj yuav tsum muab pov tseg thiab tsis noj.
6. Cov dej ntauwd yuav tsum tau ntxuav thiab tua kab mob ntau zaus.
7. Haus dej thaum mus ncig lossis mus pw hav zoov yuav tsum tau muab rhaub thiab tua kab mob.
8. Yog tias koj muaj cov tsos mob xws li ntuav, raws plab lossis ua npaws, koj yuav tsum mus ntsib kws kho mob sai li sai tau.

Kev kuaj Salmonella

Antibody kuaj

Qhov tshwj xeeb tseem ceeb ntawm cov tshuaj tiv thaiv antigen-antibody tau siv rau kev txheeb xyuas cov kab mob thiab kev ntaus ntawv serological rau ntau tshaj li ib nrab xyoo pua. Lub xub ntiag ntawm cov tshuaj tiv thaiv tshwj xeeb rau cov kab mob hauv cov kab mob los yog flagellar antigens tso cai rau tib neeg los tsim txoj hauv kev sai los xyuas cov kab mob hauv cov zaub mov. Muaj ntau txoj hauv kev los tiv thaiv kev tiv thaiv kab mob Salmonella, uas tuaj yeem muab faib ua cov txheej txheem raws li enzyme-labeled antibodies (ELISA), fluorescent antibody staining (immunofluorescence), isotope-labeled antibodies (radioimmunoassay), thiab ntau lwm txoj kev. Antibody-based txoj kev siv latex agglutination, immunosensors, immunodiffusion thiab immunochromatography. Txawm li cas los xij, txoj kev siv dav tshaj plaws yog ua raws li ob-site ELISA cov txheej txheem, uas yog cov qhaub cij ELISA. Tom qab kev txhim kho ntawm txoj kev no, radioactive isotopes yog siv los hloov cov tshuaj tiv thaiv kab mob. Hauv cov ntsiab lus, nws yog hais txog kev siv cov tshuaj "capture" antibodies immobilized ntawm cov khoom substrate los ntes cov phiaj antigen. Tom qab ntxuav kom tshem tawm cov khoom tsis sib xws, daim ntawv lo thib ob enzyme ntxiv Antibodies, uas yog khi rau ntau qhov chaw ntawm cov antigen ntes, ntxiv rau cov enzyme substrate tom qab ntxuav thib ob, thiab tso cai rau cov tshuaj tiv thaiv nrog cov xim, thiab tom qab ntawd lub hom phiaj. Antigen tuaj yeem tshawb pom tau yooj yim los ntawm spectrophotometric. Siv microtiter daim hlau raws li ib tug khoom matrix standardizes cov tshuaj tiv thaiv hom thiab pab txhawb nws automation.

Txoj kev nucleic acid

Cellular nucleic acid DNA thiab RNA tsuas yog cov molecules loj uas tuaj yeem nqa cov ntaub ntawv. Txij li txhua lub hlwb muaj cov molecule no, nws tuaj yeem siv los ua lub hom phiaj rau kev tshawb nrhiav. Lub hom phiaj feem ntau yog ib qho tshwj xeeb nucleic acid ib ntus, uas tuaj yeem tshawb pom los ntawm kev siv cov nucleic acid molecules ntxiv raws li kev sojntsuam. Zoo ib yam li cov txheej txheem tiv thaiv kab mob, qhov kev sojntsuam kuj yuav tsum tau sau kom tsim nyog, xws li xov tooj cua isotopes, enzymes lossis cov cim luminescent. Fitts et al. tau qhia thawj tiam ntawm DNA-RNA hybridization thev naus laus zis hauv kev tshawb nrhiav Salmonella hauv cov zaub mov. Cov kev sojntsuam siv nyob rau hauv txoj kev no muaj cov DNA tawg ntawm Salmonella typhi uas tau sau nrog cov isotopes radioactive. Lawv rhiab heev. Tom qab kwv yees li 48h ntawm cov kauj ruam ntxiv, qhov kev kuaj pom muaj peev xwm ncav cuag 108 kab mob / ml, tab sis vim yog siv cov xov tooj cua, uas tsuas yog siv tau rau hauv cov chaw kuaj mob tshwj xeeb, qhov zoo ntawm txoj kev no yog offset. Txog rau qhov kawg no, kev siv tshuab thib ob raws li nucleic acid hybridization, colorimeter, tau tsim.