UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

MTAALA WA UICC WA

HPV na SARATANI YA MLANGO WA KIZAZI

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Sura ya 2.d.

Uchunguzi na ubainishaji uchambuzi na uainishaji wa HPV

Prof. Suzanne Garland MD

Mkurugenzi wa Utafiti wa Maikrobiolojia, Mkuu wa Maikrobiolojia ya Kimatibabu na Magonjwa ya Kuambukiza Hospitali ya Wanawake ya Royal Melbourne, Australia Prof. Msaidizi Sepehr Tabrizi Mwanasayansi Mwandamizi wa Utafiti, Idara ya Maikrobiolojia Hospitali ya Wanawake ya Royal Melbourne, Australia UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Chembe zinazofanana na virusi (VLP)

• Analogi zisizoambukiza za kirusi kilicho na vimelea vya ugonjwa • VLP-L1 hukusanyanyika katika mandhari ya utafiti (

in vitro

) kwenye kapsidi (ganda la protini la kirusi) tupu au ‘VLP’ • Uzalishaji wa VLP hujasanifishwa • Mifumo tofauti ya maelezo, mbinu tayarishi, mitazamo ya kudhibiti ubora - Maumbo ya kutambulisha hutofautiana (moja kwa moja dhidi ya yasiyokuwa moja kwa moja) - Hakuna kiwango cha ubora cha dhahabu (gold standard) cha kuweka kizingiti cha juu cha matokeo chanya - Milinganisho michache baina ya maabara UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Chembe zinazofanana na virusi (VLP)

• VLP zinaweza kuchambulilwa kwa kutumia mbinu za hadubini ya elektroni W UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Ugunduzi wa zindiko za HPV

• Zindiko - Uwepo unaashiria maambukizi yaliyopita au yaliyopo - Zinaweza kuzalishwa dhidi ya protini ya HPV ya mapema (E) au ya kuchelewa (L) - Zindiko za kubatilisha L1 huwiana na ulinzi uliyoanzishwa kwa chanjo UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Ugunduzi wa zindiko za HPV

• Mbinu kadhaa za uchambuzi wa vijenzi (assay) zilizokuzwa ili kugundua zindiko kwenye maji maji ya damu/utando telezi: Uchambuzi wa usharabu wa kuunganisha vimeng’enya (Enzyme-linking immunosorbent assay) (ELISA): hupima jumla ya Ig, batilishi na isiyo batilishi - Chambuzi za unganishi shindani (Competitive binding assays), k.m. competitive Luminex assay (cLIA): hatua za kubatilisha Ig hadi kwa epitopi moja tu, lakini kwa vikundi vyote vya Ig - Uchambuzi wa Pseudovirion (Pseudovirion assay): hupima jumla ya ubatili wa Ig, vikundi vidogo vyote - Mbinu ya Western blot - Mbinu ya Radio-immunoprecipitation UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Ugunduzi wa DNA ya HPV katika tishu kwa kutumia hadubini

• Imunohistokemia (Immunohistochemistry) – antijeni iliyo maalum kwa kundi (group-specific antigen) • Ukaguzi wa Pap (Pap smear) : hugundua seli zinazotangulia saratani UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Mbinu za Ugunduzi wa Molekuli za DNA ya HPV

Mbinu za ukuzaji (Amplification method)

• Ukuzaji wa ishara - Hybrid Capture (HC2) • Ukuzaji wa lengo - Mjibizano wa polimerasi (Polymerase chain reaction) – Mjibizano wa polimerasi wa DNA ya HPV (PCR HPV DNA) - mRNA ya HPV (HPV mRNA) UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Uchambuzi wa vijenzi wa aina ya Hybrid Capture 2 ( Hybrid Capture 2 assay)

• Upimaji wa sasa ulioidhinishwa na FDA - Umbo la neli la1995; umbo la aina ya micro-titre la 1999 • Umesanifiwa kufanya kazi kwa kutumia sampuli ya seli zilizotolewa kwenye mlango wa kizazi • Zana za kukusanya zilizopendekezwa ni pamoja na burashi na chombo cha usafirishaji wa sampuli - Hukusanya seli za ndani na nje ya mlango wa kizazi UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Uchambuzi wa Hybrid Capture 2 assay

• Kiwango cha hisi cha 5000 jenomu/sampuli • Mseto wa uchunguzi wa kiwango cha chini cha hatari - Aina za HPV 6, 11, 42, 43, 44 • Mseto wa uchunguzi wa kiwango cha juu cha hatari - Aina za HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68 • Matokeo si maalum kwa aina fulani • Hakuna udhibiti kwa idadi ya seli zinazoingizwa • Ulinganishi mzuri baina ya maabara UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Mjibizano wa Polimerasi

• Kiwango cha juu mno cha hisi takriban nakala 1-10 /seli 100,000 • Huruhusu upimaji wa sampuli za aina kadhaa • Huruhusu upimaji wa sampuli za hifadhi zenye DNA duni zaidi • Chambuzi za maafikiano: kwa kawaida hulenga

Eneo la L1

- Huhifadhiwa - Ncha (Primers) hutofautiana MY09/MY11 SPF1/SPF2 PGMY09/PGMY11 GP5+/GP6+ - Mifumo ya Ugunduzi hutofautiana: ELISA kitone cha doa (dot blot) UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Taratibu tofauti za PCR

• Upimaji wa AMPLICOR ® HPV • Upimaji wa LINEAR ARRAY® HPV • Upimaji wa Kuainisha Jenomu wa INNO-LiPA ® - Uainishaji wa jenomu za HPV kwenye sehemu zilizotiwa kwenye mafuta ya taa - Huenda sehemu za tishu zikawa kongwe kwa kiasi fulani na zenye DNA iliyoharibika (ikiwemo mnamo wakati wa kuzidua) UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Taratibu tofauti za PCR

• Umbo la Kibanzi cha DNA ya HPV (HPV DNA Chip format) • Mkondo mdogo wa HPV (HPV Microarray) • Mkondo wa Flow Cytometry (Flow Cytometry Array) UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Taratibu nyinginezo za kugundua aina za HPV

• Luminex’s xMAP® Technology • Norchip - HPV Proofer UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Upungufu na changamoto

• Chambuzi za namna mbalimbali • Tofauti za kiwango cha hisi na za umaalum • Ukosefu wa dhibiti zinazopatikana kwa upesi, QA & QC UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Matumizi tendaji ya mbinu za kupima DNA ya HPV

Ya Utibabu

• Uchunguzi • Kuainisha matokeo yenye shaka/yasiyo dhahiri ya ukaguzi wa Pap • Kufuatilia matibabu yayofuata kugunduliwa kwa mabadiliko ya seli (post-dysplasia treatment)

Elimu ya magonjwa ya mlipuko (epidemiolojia)

• Upelelezi wa maeneo ya kijiografia - Kutapakaa kwa HPV - Kutapakaa kwa jenotipu ya HPV

Chanjo

• Utekelezaji wa kabla na baada ya chanjo - Ustadi wa majaribio ya chanjo - Maeneo tofauti ulimwenguni UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi

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Asante

Wasilisho hili linapatikana katika www.uicc.org/curriculum

UICC HPV and Cervical Cancer Curriculum Chapter 2.d. Screening and diagnosis - HPV analysis and typing Prof. Suzanne Garland, MD; Prof. Sepehr Tabrizi