Kiti Yodziwira Kusintha kwa Ma EGFR Gene 29 ya Munthu (Fluorescence PCR)-RUO

Kufotokozera Kwachidule:

[Khodi Yamalonda] HWTS- TM4008

Kufotokozera Kwachidule

Kiti iyi imagwiritsidwa ntchito kuzindikira kusintha kwa majini komwe kumachitika m'thupi la munthu pogwiritsa ntchito njira ya vitro (exons 18-21) mu jini ya EGFR m'zitsanzo kuchokera kwa odwala khansa ya m'mapapo omwe si ang'onoang'ono (onani Gome 1 kuti mudziwe zambiri).

Kugwiritsa Ntchito Kafukufuku Kokha


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Kiti Yodziwira Kusintha kwa Ma EGFR Gene 29 ya Munthu (Fluorescence PCR)-RUO

Epidemiology

Khansa ya m'mapapo yakhala chifukwa chachikulu cha imfa za khansa padziko lonse lapansi, zomwe zikuwopseza kwambiri thanzi la anthu. Khansa ya m'mapapo yomwe si ya maselo ang'onoang'ono imakhudza pafupifupi 80%-85% ya odwala khansa ya m'mapapo. Pakadali pano EGFR ndiye cholinga chachikulu cha mamolekyulu pochiza khansa ya m'mapapo yomwe si ya maselo ang'onoang'ono. Kuchuluka kwa phosphorylation ya EGFR kungathandize kukula kwa maselo a chotupa, kusiyanitsa, kulowa, kufalikira kwa maselo, komanso kulimbikitsa angiogenesis ya chotupa. EGFR tyrosine kinase inhibitors (TKI) imatha kuletsa njira yolumikizirana ya EGFR poletsa EGFR autophosphorylation, motero kuletsa kuchulukana ndi kusiyanitsa kwa maselo a chotupa, kulimbikitsa apoptosis ya maselo a chotupa, kuchepetsa angiogenesis ya chotupa, ndi zina zotero, kuti akwaniritse chithandizo cholunjika ku chotupa [1]. Kafukufuku wambiri wasonyeza kuti mphamvu yochizira ya EGFR-TKI ikugwirizana kwambiri ndi momwe EGFR gene mutation ilili, ndipo imatha kuletsa kukula kwa maselo a chotupa ndi kusintha kwa majini a EGFR [2,3]. Jini ya EGFR ili pa mkono waufupi wa chromosome 7 (7p12), ndi kutalika konse kwa 200Kb ndipo ili ndi ma exons 28. Dera la kusintha kwa majini limapezeka makamaka mu ma exons 18 mpaka 21, ma codons 746 mpaka 753 deletion mutation pa exon 19 amakhala pafupifupi 45% ndipo kusintha kwa L858R pa exon 21 kumakhala pafupifupi 40% mpaka 45% [4]. Malangizo a NCCN Okhudza Kuzindikira ndi Kuchiza Khansa ya M'mapapo Yosakhala Yaing'ono ya Maselo Ang'onoang'ono amanena momveka bwino kuti kuyesa kusintha kwa majini a EGFR kumafunika EGFR-TKI isanayambe kuperekedwa. Kiti yoyeserayi imagwiritsidwa ntchito kutsogolera kupereka mankhwala a epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), ndikupereka maziko a mankhwala apadera kwa odwala omwe ali ndi khansa ya m'mapapo yosakhala yaing'ono ya maselo. Kitiyi imagwiritsidwa ntchito kokha pozindikira kusintha kwa majini a EGFR mwa odwala omwe ali ndi khansa ya m'mapapo yosakhala ya maselo ang'onoang'ono. Zotsatira za mayesowa ndi za chithandizo chamankhwala chokha ndipo siziyenera kugwiritsidwa ntchito ngati maziko okha a chithandizo cha odwala payekha. Madokotala ayenera kuganizira za momwe wodwalayo alili, zizindikiro za mankhwala, ndi chithandizo. Zomwe zimachitika komanso zizindikiro zina zoyesera za labotale ndi zina zimagwiritsidwa ntchito poweruza mokwanira zotsatira za mayeso.

Njira

Chida Chothandizira Kuyankha Njira
EGFR IC Reaction Buffer Njira ya FAM
L858R Reaction Buffer Njira ya FAM
19del Reaction Buffer Njira ya FAM
T790M Reaction Buffer Njira ya FAM
G719X Reaction Buffer Njira ya FAM
3Ins20 Reaction Buffer Njira ya FAM
L861Q Reaction Buffer Njira ya FAM
S768I Reaction Buffer Njira ya FAM

Magawo aukadaulo

Malo Osungirako Madzi: ≤ -18℃ Mumdima
Nthawi yokhalitsa Miyezi 12
Mtundu wa Chitsanzo minofu yatsopano ya chotupa, gawo lozizira la matenda, minofu kapena gawo la matenda lomwe laphatikizidwa ndi parafini, plasma kapena seramu
CV <5.0%
LoD Kuzindikira njira yothetsera vuto la nucleic acid pansi pa 3ng/μL wild-type, kumatha kuzindikira bwino kusintha kwa 1%;

b) reaction ya nucleic acid Pansi pa 1% mutation rate, 1×103Copies/mL mutation ikhoza kupezeka mokhazikika kumbuyo kwa wild-type kwa 1×105Copies/mL;

c) Yankho la mkati mwa reference reaction limatha kuzindikira chinthu chofotokozera cha SW3 chokhala ndi malire otsika kwambiri ozindikira a internal reference ya enterprise. Pamene kit yazindikira zinthu zofotokozera za dziko lonse, zinthu zofotokozera za 2.5% mutation frequency detection ndi EGFR gene amplification mutation yokhala ndi copy number variation ya 4 zitha kuzindikirika bwino.

Kufotokozera Mwapadera Palibe njira yolumikizirana ndi DNA ya genomic ya munthu wamtundu wakuthengo ndi mitundu ina yosinthika
Zida Zogwiritsidwa Ntchito Machitidwe a Biosystems Ogwiritsidwa Ntchito 7500 Real-Time PCR,

Machitidwe a Biosystems Ogwiritsidwa Ntchito 7300 Real-Time PCR,

Makina a PCR a QuantStudio® 5 a Real-Time,

Dongosolo la LightCycler® 480 Real-Time PCR,

Dongosolo la BioRad CFX96 Real-Time PCR.

 

Kuyenda kwa Ntchito

Kiti Yodziwira Kusintha kwa Maonekedwe a Gene 29 ya Munthu (Fluorescence PCR)-RUO (2)

 

Ma reagents ofunikira koma osaperekedwa:DNase/RNase Free Water, ethanol yosalowa madzi. Poyesa chitsanzo cha minofu yolumikizidwa ndi paraffin, tikulimbikitsidwa kugwiritsa ntchito QIAamp DNA FFPE Tissue Kit (56404) yopangidwa ndi QIAGEN ndi Paraffin-embedded Tissue DNA Rapid Extraction Kit (DP330) yopangidwa ndi Tiangen Biotech (Beijing) Co., Ltd.. Poyesa chitsanzo cha plasma/serum, tikulimbikitsidwa kugwiritsa ntchito Macro & Micro-Test General DNA/RNA Kit (HWTS-3019-50, HWTS-3019-32, HWTS-3019-48, HWTS-3019-96) (yomwe ingagwiritsidwe ntchito ndi Macro & Micro-Test Automatic Nucleic Acid Extractor (HWTS-3006C, HWTS-3006B)) yopangidwa ndi Jiangsu Macro & Micro-Test Med-Tech Co., Ltd. kapena QIAamp Circulating Nucleic Acid Kit (55114) yopangidwa ndi QIAGEN.

Zogwiritsidwa ntchito ndizofunikira koma sizinaperekedwe: Malangizo a DNase/RNase Free, magolovesi otayidwa, chubu cha DNase/RNase Free Centrifuge, mipiringidzo ya machubu 8 a PCR, centrifuge.


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