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Abstrak :
ABSTRAK
Kasus infeksi measles baik individu maupun kejadian luar biasa KLB di Indonesia masih banyak ditemui. Konfirmasi infeksi measles klinis hanya dapat dilakukan di Laboratorium Nasional. Keterbatasan kit komersial yang rutin digunakan menyebabkan pemeriksaan menjadi terhambat. Pengembangan in house plate coating spesifik IgM measles dengan indirect ELISA dilakukan dengan memodifikasi dan mengoptimasi microtiter plate dengan kultur virus measles. Kultur virus measles didapat dengan menumbuhkannya pada kultur sel vero/hSLAM. Optimasi plate coating dilakukan menggunakan kultur virus measles dengan isolat MO/38/V/07 dan J/10/358/Riau dalam pengenceran 1:1 mdash;1:2.048 dan inaktif atau tidaknya virus pada saat coating dilakukan. Optimasi pemeriksaan indirect ELISA untuk in house plate coating dilakukan dengan konsentrasi konjugat 1:10, 1:25, dan 1:50. In house plate coating telah dioptimasi dan menunjukkan hasil optimum untuk mendeteksi IgM measles pada pengenceran 1:16 dengan isolat MO/38/V/07 dalam keadaan inaktif dan pemeriksaan menggunakan konsentrasi konjugat 1:25.

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ABSTRAK
Cases of infection measles both individuals and extraordinary events outbreak in Indonesia are still widely encountered. Confirmation of measles clinical infection can only be done at the National Laboratory. The limitations of commercial kits that are routinely used cause the examination to be inhibited. The development of a specific IgM measles in house plate coating with indirect ELISA is done by modifying and optimizing the microtiter plate with measles virus culture. Viral culture measles obtained by growing it on cell culture vero hSLAM. Optimization of plate coating was done using culture of measles virus with MO 38 V 07 and J 10 358 Riau isolates in dilution of 1 1 mdash 1.2048 and inactivation or active of virus at the time of coating. Optimization of indirect ELISA examination for in house plate coating is done with conjugate concentration 1 10, 1 25, and 1 50. In house plate coating was optimized and showed optimum results for detecting IgM measles at 1 16 dilution with MO 38 V 07 isolates in inactivation and examination indirect ELISA using a 1 25 conjugate concentration.

Abstrak :
Diabetes melitus merupakan kelainan metabolik yang ditandai hiperglikemia dimana landasan terapinya masih menggunakan human insulin. Telah banyak sediaan yang beredar, namun metode analisis yang standar dan valid belum ditetapkan di dalam negeri. Penelitian ini bertujuan untuk mengembangkan dan memvalidasi metode analisis secara indirect ELISA dibandingkan KCKT UV fase terbalik untuk determinasi sediaan. Antibodi poliklonal IgG dihasilkan dari kelinci yang diimunisasi dengan 1 mg/mL antigen human insulin rekombinan, dimurnikan melalui presipitasi dan kromatografi afinitas, dikuantitasi dengan spektrofotometer UV280nm, dikarakterisasi dengan uji Dot blot menggunakan substrat BCIP-NBT, serta dikarakterisasi melalui uji SDS-PAGE dan Western Blot dengan konsentrasi gel 7,5% dan 17,5%. Sedangkan, metode KCKT menggunakan kolom ReliantTM C-18 (4,6 x 150 mm, 5 µm), fase gerak Na2SO4 pH 2,3 : Na2SO4 pH 2,3 dalam asetonitril (55:45,v/v) rasio 38:62 v/v, standar internal etilparaben 10 µg/mL, detektor UV 215 nm, suhu kolom 40°C, laju alir 1 mL/menit, dan volume injek 20 µL. Validasi kedua metode dilakukan terhadap larutan uji yang mengandung m-kresol dan gliserol. Metode ELISA pada rentang 80,11-200,28 µg/mL (r = 0,99) dan KCKT pada 9,735-146,025 µg/ml (r = 0,9997) terbukti linear. Rekoveri pada ELISA dan KCKT adalah 99,11% ± 5,01 dan 100,71% ± 1,11, sedangkan RSD 3,91% dan 0,64%. LOD dan LOQ metode ELISA 22,05 µg/mL dan 73,51 µg/mL, serta KCKT 0,193 µg/ml dan 0,643 µg/ml. Human insulin bersifat stabil pada suhu 2-8°C selama 24 jam (ELISA) dan suhu 23°C selama 48 jam (KCKT). Kesimpulan, hasil validasi kedua metode valid dan mampu mendeterminasi human insulin tanpa berbeda siginifikan (Uji T, a0,05).  ......Cannot connect to Ginger Check your internet connection or reload the browser. Disable in this text field Rephrase Rephrase current sentence Edit in Ginger Enable Ginger Cannot connect to Ginger Check your internet connection or reload the browser Disable in this text field phrase. Rephrase current sentence. Edit in Ginger Enable Ginger. Diabetes mellitus is a metabolic disorder characterized by hyperglycemia that is still treated with human insulin. Many preparations on the market, but a standard and valid analytical method has not been established in our country. This study aims to develop and validate an indirect ELISA method for determining human insulin comparative to reverse phase UV HPLC. IgG polyclonal antibodies were produced by immunizing rabbits with 1 mg/mL recombinant human insulin antigen, purified by precipitation and affinity chromatography, quantified by UV spectrophotometer 280nm, characterized by dot blot test using BCIP-NBT substrate, and characterized by SDS-PAGE and Western Blot at 7.5% and 17.5% gel concentrations. The HPLC method was performed using a Reliant TM C-18 column (4.6 x 150 mm, 5 µm), with mobile phase Na2SO4 pH 2.3: Na2SO4 pH 2.3 in acetonitrile (55:45, v/v) ratio 38: 62 (v/v), 10 µg/mL ethylparaben internal standard, UV detector 215 nm, 40°C column temperature, 1 mL/minute flow rate, and 20 µL injection volume. The validation of both methods using test solutions containing m-cresol and glycerol. ELISA method in the range of 80.11-200.28 µg/mL (r = 0.99) and HPLC at 9.735-146.025 µg/ml (r = 0.9997) was resulted to be linear. The recovery yields on ELISA and HPLC were 99.11%±5.01 and 100.71%±1.11. RSD on ELISA and HPLC were 3.91%, and 0.64%, respectively. The LOD and LOQ of the ELISA were 22.05 µg/mL and 73.51 µg/mL, while HPLC were 0.193 µg/ml and 0.643 µg/ml. Human insulin is stable at 2-8°C for 24 hours (ELISA) and 23°C for 48 hours (HPLC). In conclusion, the validation results of both methods are valid and able to determine human insulin with no significant difference (T test, a0.05). Cannot connect to Ginger Check your internet connection or reload the browser Disable in this text field Rephrase Rephrase current sentence Edit in Ginger Enable Ginger Cannot connect to Ginger Check your internet connection or reload the browser Disable in this text field Rephrase Rephrase current sentence Edit in Ginger.