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"ABSTRAKGelatin merupakan salah satu jenis protein yang diperoleh dari kolagen alami yang terdapat dalam kulit dan tulang hewan. Gelatin yang paling banyak digunakan ialah gelatin babi. Penggunaan gelatin dengan bahan baku hewan babi pada produk makanan dan farmasi seringkali menyebabkan alergi, selain juga tidak diperbolehkan dalam agama islam, sehingga perlu dilakukan analisis kandungan didalam makanan olahan agar aman untuk dikonsumsi. Salah satu analisis yang dapat digunakan ialah metode imunosensor, salah satunya Enzyme-Linked Immunosorbent Assay ELISA yang memiliki tingkat sensitifias yang tinggi, mampu menguji sampel yang tidak murni, dan mampu mengikat secara selektif antigen yang dikehendaki. Metode ini memerlukan antibodi anti-gelatin babi untuk mendeteksi gelatin babi sehingga dilakukan produksi antibodi poliklonal antigen gelatin babi menggunakan hewan uji kelinci. Pada penelitian ini produksi antibodi poliklonal gelatin babi dilakukan dari antigen gelatin babi dengan mengekstraksi gelatin dari kulit babi. Rendemen yang dihasilkan sebesar 20,52 dan dikarakterisasi dengan spektrofotometer UV-Vis dan FTIR serta dianalisis dengan SDS PAGE dihasilkan bahwa gelatin babi memiliki 5 pita pemisahan dan gelatin dapat dijadikan sebagai antigen. Antibodi poliklonal antigen gelatin babi dapat diproduksi dengan hewan uji kelinci dan antibodi yang diperoleh pada hari ke 65 dilakukan pemurnian IgG. Antibodi yang dipurifikasi tersebut dapat dijadikan sebagai konjugat sensor pada imunosensor berbasis ELISA dengan konsentrasi IgG sebesar 0,289 mg/mL.

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ABSTRACTGelatin is one type of protein derived from natural collagen found in the skin and animal bones. The most widely used gelatin is pork gelatin. The use of gelatin with raw materials of pigs on food and pharmaceutical products often causes allergies, as well as not allowed in Islam, so it is necessary to analyze the content in processed foods to be safe for consumption. One of the analysis that can be used is immunosensor method, one of them is Enzyme Linked Immunosorbent Assay ELISA which has high sensitivity level, able to test impure samples, and able to selectively bind antigen desired. This method requires a pig anti gelatin antibody to detect pig gelatin so that the production of polyclonal antibody of pig gelatin antigen using rabbit test animals. In this study the production of polyclonal antigen pig gelatin carried out from pig gelatin antigen by extracting gelatin from pig skin. The yield of gelatin 20.52 and characterized by UV Vis and FTIR spectrophotometers and analyzed by SDS PAGE was found that pig gelatin has 5 separation bands and gelatin can be used as antigen. Polyclonal antibodies of antigen porcine gelatin can be produced with rabbit test animals and antibodies obtained on the 65th day of IgG purification. The purified antibodies can be used as conjugate sensors on immunosensors based ELISA with a concentration of IgG of 0.289 mg mL."

"Kasus aspergilosis paru kronik (APK) yang disebabkan Aspergillus sp. semakin meningkat seiring dengan meningkatnya frekuensi infeksi tuberkulosis (TB) paru sebagai faktor risiko. Diagnosis APK masih menjadi tantangan karena gejala klinis, pemeriksaan radiologi, maupun laboratorium tidak khas. Untuk menetapkan diagnosis APK diperlukan pemeriksaan laboratorium mikologi, termasuk uji serologi. Hasil pemeriksaan imuno-diffusion test (IDT) Aspergillus dengan crude antigen kurang optimal dan IgG Aspergillus ELISA menggunakan antigen galaktomanan yang termasuk antigen sel T independent sehingga tidak mendukung switching isotipe antibodi. Oleh sebab itu, diperlukan penelitian lain untuk mendapatkan prosedur yang lebih baik dalam menetapkan diagnosis APK. Tujuan penelitian ialah mengetahui pola respon IgG terhadap kombinasi empat protein 16 kD, 18-20 kD, 22 kD, dan 30 kD antigen Aspergillus dengan metode Western Blot. Potensi diagnostik dari kombinasi protein 16 kD, 18-20 kD, 22 kD, dan 30 kD antigen Aspergillus dengan metode Western Blot terhadap nilai konsensus positif dan negatif APK berdasarkan 2 metode pemeriksaan yaitu biakan jamur dan IgG anti galaktomanan sebagai baku emas didapatkan nilai sensitivitas dan spesifisitas sebesar 74% dan 96%. Deteksi IgG Aspergillus metode Western Blot menunjukkan antigen dominan berat molekul 16 kD dan 18-20 kD. Kesimpulan uji IgG Aspergillus Western Blot memiliki potensi diagnostik lebih baik dibanding uji IgG Aspergillus ELISA.

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Chronic pulmonary aspergillosis (CPA) caused by Aspergillus sp. potentially increases with the increasing frequency of pulmonary tuberculosis (TB) as a risk factor. Diagnosis of CPA is still a challenge because clinical symptoms, radiological examination, and laboratory are  not specific. The diagnosis of CPA needs to be performed by specific mycological examination, including serology test. The result of the Aspergillus immunodiffusion test (IDT) with crude antigen is sub optimal and Aspergillus IgG ELISA method uses galactomannan antigens that are independent T cell antigens, so cant support switching of isotype antibodies. Therefore, other study is needed to get a better procedure in determine the CPA diagnostic. The study aimed to determine the IgG responses with a combination of  Aspergillus proteins (16 kD, 18-20 kD, 22 kD, and 30 kD) with Western Blot method. Diagnostic potential of the Aspergillus protein combination (16 kD, 18-20 kD, 22 kD, and 30 kD) with Western Blot method on positive and negative consensus values of CPA based on two examination methods are fungus culture and anti-galactomannan IgG as gold standard are obtained sensitivity and specificity of 74% and 96%. From the Aspergillus Western Blot IgG test, dominant antigens obtained were molecular weights of 16 kD and 18-20 kD. The conclusion is Aspergillus specific IgG test with Western Blot method has better diagnostic potential than the  anti-galactomannan IgG ELISA method."

"Latar belakang: Karies merupakan salah satu penyakit kronis dalam rongga mulut dengan angka kejadian cukup tinggi pada anak-anak. Karies gigi yang terjadi pada bayi dan anak usia pra-sekolah dikenal dengan istilah Early Childhood Caries (ECC). Data penelitian dari beberapa negara Asia Tenggara menunjukkan prevalensi kejadian ECC pada anak usia 3-6 tahun sebanyak 25-95%. Selain itu, sebuah penelitian di Jakarta menunjukkan prevalensi ECC sebesar 52,7%. Etiologi ECC melibatkan interaksi antara organisme patogen, substrat karbohidrat terfermentasi, kerentanan host, dan waktu. Sebagai salah satu faktor host, saliva berperan dalam mempertahankan keseimbangan dinamis antara demineralisasi dan remineralisasi. Saliva mengandung proline-rich protein (PRP) yang memiliki sifat antibakteri. Tujuan penelitian ini adalah menganalisis kadar proline-rich protein saliva sebagai indikator Early Childhood Caries. Metode Penelitian: Desain penelitian ini adalah potong lintang analitik secara laboratorik. Penelitian ini dilakukan pada 14 anak dengan ECC dan 14 anak bebas karies. Saliva diperoleh dari seluruh subjek dan kadar PRP diukur menggunakan metode ELISA sandwich. Hasil: Analisis data menggunakan uji Mann Whitney U menunjukkan terdapat perbedaan bermakna secara statistik (p<0,05) antara kadar PRP saliva anak ECC dan anak bebas karies. Kesimpulan: Kadar proline-rich protein saliva dapat digunakan sebagai indikator Early Childhood Caries.

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Backgrounds: Caries is one of the chronic diseases in the oral cavity with a fairly high incidence in children. Caries experienced by infants and pre-school children is known as Early Childhood Caries (ECC). Research data from several countries in Southeast Asia showed that the prevalence of ECC in children aged 3-6 years old ranges from 25 to 95%. In addition, a study in Jakarta showed prevalence of Early Childhood Caries about 52.7%. Etiology of ECC involves interactions between pathogenic organisms, fermented carbohydrate substrates, host vulnerabilities, and time. As one of host factor, saliva plays a role in maintaining a dynamic balance between demineralization and remineralization. Saliva contains proline-rich protein (PRP) that has antibacterial properties. The purpose of this study was to analyze the concentration of proline-rich protein saliva as an indicator of Early Childhood Caries. Methods: The design of this study is cross-sectional analytical laboratory. This study was conducted on 14 children with ECC and 14 caries-free children. Saliva were taken from all subjects and the PRP levels were measured using ELISA sandwich method. Results: Data analysis using the Mann Whitney U test showed that there were statistically significant differences (p<0.05) between the levels of salivary proline-rich protein in children with ECC and caries-free children. Conclusion: The levels of salivary proline-rich protein can be used as an indicator of Early Childhood Caries."

"Latar belakang: Pasien asma persisten berisiko mengalami penyakit jamur paru, termasuk aspergilosis bronkopulmoner alergika (ABPA) dan aspergilosis paru kronik (APK). Deteksi IgG spesifik Aspergillus metode enzyme-linked immunosorbent assay (ELISA) merupakan pemeriksaan mikologi yang direkomendasikan untuk diagnosis APK. Metode baru yang lebih praktis adalah immunochromatography test (ICT) Aspergillus, tetapi data hasil pemeriksaan tersebut masih terbatas di Indonesia. Penelitian ini bertujuan untuk mengetahui nilai diagnostik ICT Aspergillus dibandingkan dengan metode ELISA pada pasien asma persisten.Metode: Uji diagnostik dilakukan untuk mengetahui nilai diagnostik ICT Aspergillus dibandingkan dengan IgG spesifik Aspergillus metode ELISA. Bahan klinis berasal dari serum pasien asma persisten di RSUP Persahabatan pada tahun 2021 yang memenuhi kriteria inklusi. Deteksi antibodi kedua metode tersebut dilaksanakan di laboratorium Departemen Parasitologi FKUI untuk mengetahui proporsi, sensitivitas, spesifisitas, nilai duga positif dan negatif.Hasil: Deteksi antibodi spesifik Aspergillus kedua metode dilakukan terhadap 50 serum pasien. Proporsi hasil positif ICT Aspergillus 16%, sedangkan ELISA 32%. Nilai diagnostik ICT Aspergillus dibandingkan ELISA menunjukkan sensitivitas 25%, spesifisitas 88,24%, nilai duga positif 50%, dan nilai duga negatif 71,43%.Kesimpulan: Pemeriksaan ICT Aspergillus belum dapat direkomendasikan sebagai pemeriksaan standar diagnosis APK pada pasien asma persisten karena memiliki sensitivitas rendah

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Introduction: Patients with persistent asthma are at risk of developing pulmonary mycoses, including allergic bronchopulmonary aspergillosis (ABPA) and chronic pulmonary aspergillosis (CPA). Aspergillus immunochromatography test (ICT) is a new practical method to detect Aspergillus-specific antibodies, but data on ICT results are still limited in Indonesia. This study aims to determine the diagnostic value of Aspergillus ICT compared to ELISA in persistent asthma patients.

Methods: Diagnostic tests were carried out to determine the diagnostic value of Aspergillus ICT. The antibody detection on the sera of persistent asthma patients at Persahabatan General Hospital in 2021 who met inclusion criteria was carried out in the laboratory of Parasitology Department, FMUI to determine the proportion, sensitivity, specificity, positive and negative predictive values.

Results: Detection of Aspergillus-specific antibodies was performed on 50 patient sera using ICT and ELISA. The proportion of positive results for Aspergillus ICT revealed 16%, while the ELISA 32%. The diagnostic value of Aspergillus ICT compared to ELISA showed a sensitivity of 25%, specificity of 88.24%, a positive predictive value of 50%, and a negative predictive value of 71.43%.

Conclusion: Aspergillus ICT could not be recommended as a standard examination for CPA diagnosis in persistent asthma patients due to its low sensitivity."

"Latar belakang: Aspergilosis paru kronik (APK) merupakan komplikasi yang sering ditemukan pada pasien dengan riwayat tuberkulosis (TB) paru. Pemeriksaan serologi Aspergillus yang cepat dan sensitif dibutuhkan untuk mendiagnosis APK. Penelitian ini membandingkan pemeriksaan serologi ELISA otomatis dan tes imunokromatografi pada dalam mendeteksi antibodi spesifik Aspergillus pada pasien dengan riwayat TB paru.Metode: Studi potong-lintang ini merupakan bagian dari penelitian payung tentang diagnosis APK pada pasien TB. Serum pasien diperiksa menggunakan ELISA otomatis dan tes imunokromatografi. Performa diagnosis diperoleh melalui perbandingan hasil pemeriksaan dengan diagnosis APK. Agreement antar tes dianalisis dengan Hasil: Didapatkan 68 pasien pasca TB dengan median usia 34,5 (17-72) tahun. Sebanyak 24 pasien diadiagnosis APK (35,3%). Proporsi positif ELISA otomatis dan tes imunokromatografi sebesar 48,5% dan 2,9%. ELISA otomatis memiliki sensitivitas 91,7%, spesifisitas 75%, nilai duga positif 66,7%, dan nilai duga negatif  94,3%. Tes imunokromatografi memiliki sensitivitas 8,3%, spesifisitas  dan nilai duga positif 100%, serta nilai duga negatif 66,7%. Agreement antara kedua tes sangat rendah (kappa score: 0,062).Kesimpulan: Pada pasien dengan riwayat TB paru, ELISA otomatis IgG spesifik Aspergillus dapat digunakan sebagai pemeriksaan penunjang APK di area dengan sumber daya mendukung. Sedangkan tes imunokromatografi dapat digunakan sebagai uji penapisan awal APK di daerah dengan keterbatasan sumber daya.

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Introduction: Chronic pulmonary aspergillosis (CPA) is common in post TB patients. Faster and more sensitive Aspergillus serological examination is necessary for CPA diagnosis. This study compared automated ELISA and immunochromatography test for Aspergillus specific antibody detection in post pulmonary TB patients.

Method: This cross-sectional study is part of previous research on CPA diagnosis in tuberculosis. Patient’s sera were tested with automated ELISA and immunochromatography test. The results were compared to CPA diagnosis to obtain diagnostic performances. The agreement between tests was analyzed with Cohen’s kappa.

Result: There were 68 previous TB patients with median age 34,5 (17-72) years old. CPA was diagnosed in 24 patients (35,3%). The positive result of automated ELISA and immunochromatography test were 48,5% and 2,9%. Automated ELISA showed sensitivity of 97%, specificity of 75%, PPV of 66,7%, and NPV of 94,3%. The immunochromatography test showed sensitivity of 8,3%, specificity and PPV of 100%, and NPV of 66,7%. There was very low agreement between tests (kappa score: 0,062).

Conclusion: Automated ELISA Aspergillus specific IgG could be utilized for supporting CPA diagnosis in post TB patients, mainly in settings where resources are available. Immunochromatography test is applicable as an early screening tool for CPA detection in resource-constrained areas."

"Penelitian dengue secara in vitro banyak dilakukan untuk mengetahui mekanisme pasti patogenesis infeksi degue. Namun, sedikitnya informasi mengenai karakter pertumbuhan virus dengue pada galur sel model menjadi salah satu faktor pembatas. Karakterisasi pertumbuhan kinetik dilakukan untuk mengetahui dengan melihat karakter pertumbuhan kinetik virus dengue pada galur sel C6/36, Vero76, MDCK, 293, HepG2, dan A549. Parameter pertumbuhan virus dengue pada tiap sel diketahui dengan melihat kecepatan replikasi, ekspresi protein NS1, dan deteksi genom virus dengue. Hasil uji kinetik menunjukkan adanya perbedaan profil pertumbuhan tiap serotipe virus dengue pada tiap galur sel, dengan pertumbuhan relatif lebih tinggi pada sel A549 dibandingkan dengan galur sel lainnya. Hasil uji ELISA menunjukkan ekspresi protein NS1 pada sel A549 meningkat seiring peningkatan titer virus. Keberadaan RNA genom virus dengue pada sel A549 dikonfirmasi menggunakan RT-PCR. Dengan demikian, keseluruhan hasil penelitian ini menunjukkan virus dengue mampu tumbuh dengan baik pada galur sel A549, sehingga dapat digunakan sebagai galur sel mamalia alternatif untuk propagasi dan penelitian infeksi virus dengue lebih lanjut.

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In vitro dengue research has been routinely used to study the pathogenesis of dengue infection. The little information about the growth characteristics of dengue viruses in various cell lines model has become the limiting factor of dengue research. Dengue virus growth characterization was conducted to determine the growth kinetics of dengue viruses in several cell lines i.e. in C6/36, Vero76, MDCK, 293, HepG2, and A549 cell lines. Growth characteristics of dengue virus in each cell were measured by looking at the rate of replication, NS1 protein expression, and detection of dengue virus genome. The replication kinetic assay indicated the difference growth characteristics of each serotype of dengue virus in each cell line, with the relatively higher growth was observed in A549 cell line compared to other cell lines. ELISA result showed an increased expression of NS1 protein in A549 cell in parallel with increasing viral titer. The presence of dengue virus RNA genome in A549 cells was confirmed using RTPCR assay. This study observed the ability of dengue viruses to grow well in A549 cell line and this cell line could be used as an alternative mammalian cell line for propagation and further study of dengue virus infection."

"Masih tingginya prevalensi infeksi HIV di Indonesia dan anjuran WHO untuk melakukan pengembangan vaksin HIV-1 dalam penanganan penyebaran infeksi, menjadikan pentingnya dilakukan pengembangan vaksin HIV untuk tujuan preventif maupun kuratif HIV-1 berdasarkan subtipe AE_CRF01 isolat Indonesia. MPER gp41 sebagai salah satu target dalam pengembangan vaksin HIV telah diketahui dapat menginduksi produksi antibodi netralisasi HIV terhadap MPERgp41. Namun dalam implementasinya, MPER memiliki imunogenisitas yang rendah. Pada penelitian terdahulu telah berhasil dibuat vaksin DNA HA-MPER2 yang mengandung gen HA H5N1 sebagai antigen yang akan mempresentasikan epitop MPER gp41 HIV-1. Penelitian ini dilakukan untuk membuktikan adanya respon antibodi spesifik MPER gp41 HIV-1 pada mencit BALB/c yang diimunisasi vaksin DNA HA-MPER2, kemudian dibandingkan dengan kelompok mencit yang diimunisasi vaksin DNA HA-MPER1, vaksin DNA HA dan vaksin DNA wildtype. Penelitian diawali dengan membuat sel E. coli TOP 10 kompeten, transformasi plasmid rekombinan, verifikasi hasil isolasi plasmid dengan analisa gel agarosa 0,8% dan sekuensing. Kemudian dilakukan preparasi vaksin DNA sebelum penyuntikan dengan dilarutkannya plasmid dalam PBS 1x dan ditambahkan DMRIE-c (perbandingan molar 1:2). Pengujian ELISA dilakukan terhadap serum antibodi (pengenceran 1/50) dengan menggunakan peptida ELDKWAS 12,5 μg/ml sebagai antigen. Hasil uji ELISA menunjukkan adanya respon antibodi spesifik terhadap MPER gp41 HIV-1 dengan titer antibodi dalam jumlah rendah.

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The high prevalence of HIV infection in Indonesia and WHO recommendation to develop HIV-1 vaccine in the handling of infection spreading signifies the importance of HIV-1 vaccine development for preventive and curative purpose based on AE_CRF01 subtype of Indonesian isolate. MPER gp41 as one of the target of HIV vaccine development had been known to induce production of neutralizing antibody against HIV MPER gp41. However, in the implementation, MPER had low immunogenicity. Previous work on HIV vaccine development at IHVCB-UI had succesfully produced DNA HA-MPER2 vaccine prototype which contained HA H5N1 gen as a scaffold that would present MPER gp41 HIV-1 epitope. This research was conducted to prove the existence of specific antibody response towards HIV MPER gp41 in BALB/c mice immunized by HA-MPER2 DNA vaccine in comparison with BALB/c mice immunized by HA DNA vaccine and the pcDNA3.1 vector DNA. The research was initiated by generation of E. coli TOP 10 competent cells, followed by transformation of competent cells, and plasmid isolation. The result would be verified by 0.8% agarosa gel analysis and sequencing, followed by DNA vaccine preparation by dissolving plasmid in PBS 1x and adding DMRIE-c (molar comparation 1:2). The ELISA test was conducted towards antibody serum (dilution 1/50) by using 12.5 μg/ml ELDKWAS peptide as an antigen. The result showed the presence of specific antibody response towards HIV MPER (ELDKWAS) gp41 that was observed in low titer."

"Peningkatan konsentrasi kristal monosodium urea pada sendi dan jaringan menunjukkan adanya keradangan Gout artritis (GA). Angka insiden dan prevalensi GA tersebar di negara berkembang sebesar 2?15%. Di Indonesia, prevalensi GA sekitar 29% dan sering terjadi pada suku Minahasa, Toraja dan Batak. Penelitian ini bertujuan untuk menganalisis aplikasi minyak atsiri kunyit sebagai anti-radang pada penderita GA dengan diet tinggi purin serta mengukur mediatorseluler tumor necrosis factor-α (TNF-α). Desain penelitian adalah randomized pretest-posttest control group design dengan pemberian secara single blind. Tes GCMS dilakukan untuk mengetahui komponen aktif minyak atsiri. Sampel penelitian ini adalah pasien baru GA di RS Haji Surabaya. Selama tujuh hari, kelompok perlakuan diberi minyak atsiri kunyit dengan dosis 25 mg/kg BB, sedangkan kelompok kontrol diberi indometasin dengan dosis 150 mg/kg BB. Sampel darah diambil sebelum dan sesudah perlakuan. Minyak atsiri kunyit mempunyai empat fraksi komponen aktif. Terdapat penurunan kadar urea darah pada kelompok perlakuan (p = 0,001) dan kelompok kontrol (p = 0,007). Terdapat penurunan konsentrasi pelepasan TNF-α, tetapi penurunan ini tidak berbeda secara signifikan pada kelompok kontrol dan perlakuan.Increased concentrations of crystal monosodium urea at joint and soft tissue represent induced of inflammation at gout arthritis (GA). Incidence and prevalence GA disseminate wide in developed countries in Asian range from 2-15% and In Indonesia, GA prevalence was 29% and mostly found in Minahasa, Toraja, and Batak ethnics. This research was aimed to analyse application of curcuma domestica volatile oil as anti inflammation agent on gout arthritis patient who has high purin diet and to assess specific cellular mediator Tumor Necrosis Factor-α (TNF-α). The design of the study was randomized pretest-posttest control group design with single blind treatment. The GCMS test was performed to identify active component in volatile oil. The sample was the new gout arthritis patient in Haji Public Hospital Surabaya. For a week, treatment group was assigned with volatile oil with dose 25 mg/kg body weight and the control group was given indometasin 150 mg/kg body weight. Blood samples were taken before and after treatment. Volatile oil of curcuma domestica (Curcuma domestica, val) has four fraction of active component. There wasdecreasing in blood urea level in treatment group (p = 0.001) and control group (p = 0.007). Both in control and treatment group, there was also decreasing in TNF-α, however it was not statistically significant. "

"ABSTRAKArbovirus (arthropode-borne virus) yang timbul dan timbul kembali telah memengaruhi berbagai aspek kehidupan manusia. Infeksi arbovirus terbanyak di Indonesia: dengue, Japanese encephalitis (JE) dan chikungunya (CHIK) menyebabkan kasus luar biasa tiap tahun. Ketersediaan metode deteksi JE dan CHIK sangat terbatas di Indonesia. Pengembangan in-house IgM antibody-capture Enzyme Linked Immunosorbent Assay (MAC ELISA) dengan antigen local terinaktivasi akan meningkatkan deteksi dan pemantauan dengan meningkatkan spesifisitas dan sensitivitas. Antigen diproduksi dalam kultur sel dengan sel BHK-21 dan sel Vero kemudian diinaktivasi dengan gamma-irradiasi dan 0,01% beta-propiolakton. Kinerja Antigen dievaluasi dengan uji MAC ELISA dan titer virus dihitung dengan uji plak. Virus Japanese encephalitis dan chikungunya terinaktivasi pada 20 kGy gamma- irradiasi dan 0,01% BPL. In-house MAC ELISA telah dioptimisasi dengan inkubasi 2 jam. Kit in-house MAC ELISA yang telah dikembangkan berguna untuk deteksi dan pemantauan JE dan Chik dengan fasilitas terbatas.

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ABSTRACTThe emerging and re-emerging arthropod-borne viruses (arboviruses) have effected many aspects of human existence. Three major arbovirus infection in Indonesia: dengue, Japanese encephalitis (JE) and chikungunya (CHIK) causes numerous outbreaks each year. However, availability of detection methods for JE and CHIK are very limited in Indonesia. Development of in-house IgM antibody-capture Enzyme Linked Immunosorbent Assay (MAC ELISA) with inactivated local antigen will improve detection and surveillance capability across Indonesia by increasing its specificity and sensitivity. Antigens were produced in cell culture using BHK-21 cells and Vero cells then inactivated using gamma-irradiation and 0.01% beta-propiolactone (BPL). Antigen performance was evaluated using MAC ELISA and virus titer were calculated using plaque assay. Japanese encephalitis virus and chikungunya virus was inactivated at 20 kGy with 0.01% BPL. Optimized in-house MAC ELISA protocol using these antigen has been developed. Developed in-house MAC ELISA kit will be beneficial for detection and surveillance of JE and CHIK with limited facility. "

"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). 

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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."