Pseudorabies virus (gB) PCR ukuthola ikhithi

Incazelo emfushane:

Leli khithi lisebenzisa indlela ye-PCR ye-fluorescent yesikhathi sangempela ukuthola i-RNA ye-Pseudorabies virus (gB gene) (PRV) ezintweni zezifo zezicubu ezifana namathoni, ama-lymph node kanye ne-spleen kanye nezifo zezifo ezinjenge-vaccine negazi lezingulube.


Imininingwane yomkhiqizo

Amathegi womkhiqizo

Igama lomkhiqizo

I-Pseudorabies virus (gB) I-PCR Detection Kit (iLyophilized)

Usayizi

Izivivinyo ezingama-48 / ikhithi, ama-50tests / kit

Ukusetshenziswa okuhlosiwe

Leli khithi lisebenzisa indlela ye-PCR ye-fluorescent yesikhathi sangempela ukuthola i-RNA ye-Pseudorabies virus (gB gene) (PRV) ezintweni zezifo zezicubu ezifana namathoni, ama-lymph node kanye ne-spleen kanye nezifo zezifo ezinjenge-vaccine negazi lezingulube. Kukufanele ukutholakala, ukuxilongwa kanye nophenyo lwezifo ze-Pseudorabies virus (gB gene). Ikhithi iyi-ALL-READY PCR SYSTEM (Lyophilized), equkethe i-enzyme ye-DNA amplification, reaction buffer, primers ethize kanye nama-probes adingekayo ukuthola ukutholwa kwe-RT-PCR ye-fluorescent.

Okuqukethwe komkhiqizo

Izingxenye Iphakheji incazelo Isithako
Umxube we-PRV PCR 1 × ibhodlela (i-Lyophilized powder)  50Ukuhlolwa Ama-dNTPs, ama-MgCl2, AmaPrimers, AmaProbes, iTaq DNA polymerase
I-6 × 0.2ml 8 ithubhu yomugqa omuhle(AbakwaLyophilized) 48Ukuhlolwa
Ukulawula Okuhle 1 * 0.2ml tube (i-lyophilized)  10Uvivinyo

I-Plasmid noma i-Pseudovirus equkethe izingcezu ezithile ze-PRV (gB)

Ukuqedwa kwesisombululo 1.5 ml uCryotube 500uL /
Ukulawulwa Okubi 1.5 ml uCryotube 200uL 0.9% i-NaCl

Isitoreji & Impilo Yesheluli

(1) Ikhithi ingahanjiswa ekamelweni lokushisa.

(2) Impilo yeshalofu izinyanga eziyi-18 ngo -20-nezinyanga eziyi-12 ku-2 ℃ ~ 30 ℃.

(3) Bona ilebula elikukhithi yedethi yokukhiqiza nosuku lokuphelelwa yisikhathi.

(4) I-reagent yenguqulo ye-lyophilized powder kufanele igcinwe ku -20 ℃ ngemuva kokuqedwa kanye neqhwa eliphindiwe -thaw kufanele libe ngaphansi kwezikhathi ze-4.

Izinsimbi

I-GENECHECKER UF-150, i-UF-300 real-time fluorescence PCR instrument.

Umdwebo Wokusebenza

a) Uhlobo lwebhodlela:

1

b) Uhlobo lweshubhu olu-strip olu-8 kahle:

2

Ukukhuliswa kwe-Pcr

Isilungiselelo Esinconyiwe

Isinyathelo

Umjikelezo

Ithempelesha (℃)

Isikhathi

Isiteshi se-Fluorescence

1

1

95

2min

/

2

40

95

5s

/

60

10s

Qoqa i-FAM fluorescence

* Qaphela: Izimpawu zeziteshi ze-FAM fluorescence zizoqoqwa ngo-60 ℃.

Ukuhumusha Imiphumela Yokuhlola

Isiteshi

Ukuhunyushwa kwemiphumela

Isiteshi se-FAM

Ct≤35

I-PRV (gB) Eqondile

Undet

I-PRV (gB) inegethivu

35<Ct≤40

Suspicious resut, retest*

*If the retest result of FAM channel has a Ct value ≤40 and shows typical “S” shape amplification curve, the result is interpreted as positive, otherwise it is negative.


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