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Monkeypox Virus Ixesha lokwenyani PCR Kit

Inkcazelo emfutshane:

Ikati.Hayi B001P-01 Ubungakanani Iimvavanyo ezingama-48/Ikhithi
Umzekelo ISerum / Lesion Exudate Trans.& Sto.Temp. -25~-15℃
Ifomathi Premix yaba lyophilized kwaye aliquoted kwi 8 strip PCR iityhubhu
Isixhobo Esisebenzayo Isixhobo sePCR sexesha lokwenyani njenge ABI7500, Bio-Rad CFX96, SLAN-96S, QuantStudio kunye neBTK-8

Iinkcukacha zeMveliso

Iithegi zeMveliso

Iinkcukacha zemveliso

Ukusetyenziswa okucetywayo

Isetyenziselwa ukubhaqa i-Monkeypox Virus kwi-serum yabantu okanye iisampuli ze-lesion exudate ngokusebenzisa iinkqubo ze-PCR zexesha lokwenyani.

Umgaqo woVavanyo

Le mveliso yinkqubo yovavanyo ye-PCR esekelwe kwi-fluorescent probe-based Taqman® real-time PCR.Iiprimers ezithile kunye neeprobe ziyilelwe ukuchongwa kweF3L gene yeMonkeypox Virus.Ulawulo lwangaphakathi olujolise kwimfuza egciniweyo yomntu ebeka iliso kwingqokelela yesampulu, ukuphathwa kwesampulu kunye nenkqubo yePCR yexesha langempela ukuphepha iziphumo ezingezizo zobuxoki.Ikhithi yinkqubo epheleleyo ye-premix lyophilized, equka izinto ezifunekayo kwiMonkeypox Virus ukubona: i-nucleic acid amplification enzyme, i-enzyme ye-UDG, i-reaction buffer, i-primer ethile kunye ne-probe.

Imixholo ephambili

Amacandelo anikiweyo adweliswe kwitheyibhile.

Amacandelo

Iphakheji

Isithako

Intsholongwane yeMonkeypoxI-Lyophilized Premix Iityhubhu ezi-8 zePCR× iipowutshi ezi-6 Iiprimers, probes, dNTP/dUTP Mix, Mg2+, Taq DNA polymerase, UDG Enzyme
Ulawulo oluhle lweMPV 400 μL×1 ityhubhu Ulandelelwano lwe-DNA equlethe ijini ekujoliswe kuyo
Ulawulo olubi lweMPV 400 μL×1 ityhubhu Ulandelelwano lwe-DNA equlethe icandelo lemfuza yomntu
Ukunyibilikisa Isisombululo 1 mL × 1 ityhubhu Isizinzi
Isatifikethi sokuThobela Iqhekeza eli-1

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Operation Flow

1. IsampuliIngqokelela:Isampulu kufuneka iqokelelwe kwiityhubhu eziyinyumba ngokuhambelanayo

kunye neenkcukacha zobugcisa ezisemgangathweni.

2. ULungiselelo lwe-Reagent (iNdawo yokuLungiselela i-Reagent)

Thatha amacandelo ekhithi, uwalinganisele kwiqondo lobushushu begumbi ukwenzela ukusetyenziswa kokulinda.

3. Ukwenziwa koMfanekiso (iNdawo yokuLungiselela iSampuli)

3.1 Ukukhutshwa kwe-Nucleic acid

Kucetyiswa ukuba kuthathwe iisampulu zolwelo ezingama-200μL, Ulawulo olulungileyo kunye noLawulo olubi lwe-nucleic acid extraction, ngokweemfuno ezihambelanayo kunye neenkqubo zeekhithi zokutsalwa kwe-viral DNA.

3.2 I-Lyophilized powder yokunyibilika kunye nokudibanisa itemplate

Lungiselela iMonkeypox Virus Lyophilized premix ngokwenani leisampulu.Isampuli enye idinga ityhubhu ye-PCR ene-Lyophilized premix powder.Ulawulo olubi kunye nolawulo olulungileyo kufuneka luphathwe njengeesampuli ezimbini.

(1) Yongeza i-15μL Ukunyibilika kweSisombululo kwityhubhu nganye ye-PCR equlethe i-Lyophilized premix, uze udibanise i-5μL ekhutshiweyo iisampulu / Ulawulo olubi / uLawulo oluLungileyo kwityhubhu nganye ye-PCR ngokulandelelana.

(2) Gubungela iityhubhu ze-PCR ngokuqinileyo, nqakraza iityhubhu zePCR ngesandla de umgubo we-lyophilized unyibilike ngokupheleleyo kwaye udityaniswe, uqokelele ulwelo ukuya ezantsi kwiityhubhu zePCR ngesantya esisezantsi secentrifugation.

(3) Ukuba usebenzisa isixhobo esiqhelekileyo se-PCR yexesha eliqhelekileyo lokubona, ke udlulisele ngokuthe ngqo iityhubhu zePCR kwindawo yokukhulisa;ukuba usebenzisa i-BTK-8 ukuze ubhaqwe, ngoko kufuneka wenze le misebenzi ilandelayo: ukudlulisa i-10 μL yolwelo ukusuka kwityhubhu ye-PCR ukuya kwi-chip yokusabela kakuhle ye-BTK-8.Ityhubhu enye yePCR ihambelana nequla elinye kwitshiphu.Ngethuba lokusebenza kombhobho, qinisekisa ukuba i-pipette i-90 degrees nkqo.Iingcebiso ze-aerosol barrier pipette kufuneka zibekwe embindini womthombo ngamandla aphakathi kwaye ziyeke ukutyhala i-pipette xa ifikelela kwi-gear yokuqala (ukuphepha ama-bubbles).Emva kokuba iqula lizaliswe, thatha inwebu ye-chip yokusabela ukugquma onke amaqula kwaye itshiphu idluliselwe kwindawo yokufumanisa ukukhulisa.

4. Ukwandiswa kwePCR (iNdawo yoFundo)

4.1 Beka iityhubhu ze-PCR / i-reaction chip kwitanki yokusabela kwaye usete amagama empendulo nganye kakuhle ngokulandelelana okuhambelanayo.

4.2 Iisetingi zokufumanisa i-fluorescence: (1) iMonkeypox virus (FAM);(2) Ulawulo lwangaphakathi (CY5).

4.3 Qhuba le migaqo ilandelayo yokukhwela ibhayisekile

Iprothokholi ye-ABI7500, i-Bio-Rad CFX96, i-SLAN-96S, i-QuantStudio:

Amanyathelo

Ubushushu

Ixesha

Imijikelo

1

I-denaturation yangaphambili

95℃

2 imiz

1

2

I-Denaturation

95℃

10 s

45

I-Anealing, ukwandiswa, ukufumana i-fluorescence

60℃

30 s

 Inkqubo ye-BTK-8:

Amanyathelo

Ubushushu

Ixesha

Imijikelo

1

I-denaturation yangaphambili

95℃

2 imiz

1

2

I-Denaturation

95℃

5 s

45

I-Anealing, ukwandiswa, ukufumana i-fluorescence

60℃

14 s

5. Uhlalutyo lweziphumo (nceda ubhekisele kwi-Instrument User Manual)

Emva kokusabela, iziphumo ziya kugcinwa ngokuzenzekelayo.Cofa "Hlaziya" ukuhlalutya, kwaye isixhobo siya kutolika ngokuzenzekelayo amaxabiso e-Ct yesampuli nganye kwikholamu yesiphumo.Iziphumo zolawulo ezingalunganga nezilungileyo ziyakuhambelana noku kulandelayo "6. Ulawulo loMgangatho".

6. Ulawulo lwemeko

6.1 Ulawulo olubi: Akukho Ct okanye i-Ct> 40 kwitshaneli ye-FAM, i-Ct≤40 kwi-CY5 channel ene-curve yokukhulisa eqhelekileyo.

6.2 Ulawulo oluLungileyo: I-Ct≤35 kwitshaneli ye-FAM ene-curve ye-amplification eqhelekileyo, i-Ct≤40 kwi-CY5 channel ene-curve ye-amplification eqhelekileyo.

6.3 Iziphumo zisebenza ngokusemthethweni ukuba zonke ezi khrayitheriya zifikelelwe.Ngaphandle koko, umphumo awusebenzi.

Isiphumo sokutolika

Ezi ziphumo zilandelayo zinokwenzeka:

  Ct ixabiso letshaneli yeFAM Ixabiso le-CT ye-CY5 channel Ukutolikwa

1#

Akukho Ct okanye Ct>40

≤40

Intsholongwane yeMonkeypox ayinayo

2#

≤40

Naziphi na iziphumo

Intsholongwane yemonkeypox

3#

40-45

≤40

Uvavanyo kwakhona;ukuba isengama 40 ~ 45, xela njenge 1#

4#

Akukho Ct okanye Ct>40

Akukho Ct okanye Ct>40

Ayisebenzi

PHAWULA: Ukuba iziphumo ezingasebenziyo zenzekile, isampuli kufuneka iqokelelwe kwaye ivavanywe kwakhona.

Ulwazi lweodolo

Igama lemveliso Ikati.Hayi Ubungakanani Umzekelo Beka ubomi kwishelufa Trans.& Sto.Temp.
Monkeypox Virus Ixesha lokwenyani PCR Kit B001P-01 Iimvavanyo ezingama-48/ikhithi ISerum / Lesion Exudate Iinyanga ezili-12 -25~-15℃

  • Ngaphambili:
  • Okulandelayo:

  • Bhala umyalezo wakho apha kwaye uwuthumele kuthi