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"Background: the global scale-p of antiretroviral therapy (ART) is the primary factor contributing to the decline in deaths from acquired immune deficiency syndrome (AIDS)-related illnesses. However, the emergence of transmitted drug resistance (TDR) compromises the effects of ART in treatment-naive individuals, which may hinder treatment success. The present study aimed to identify the presence of TDR among treatment-naive individuals in Buleleng, Bali, which is currently ranked sixth among Indonesian provinces with the highest cumulative human immunodeficiency virus type 1 (HIV-1) infection cases. Methods: thirty-nine ART-naive individuals in Buleleng Regency General Hospital were enrolled in the present study. Blood samples from participants were subjected to a genotypic analysis. Results: 28 protease (PR) and 30 reverse transcriptase (RT) genes were successfully amplified and sequenced from 37 samples. HIV-1 subtyping revealed CRF01_AE as the dominant circulating recombinant form in the region. No TDR for PR inhibitors was detected; however, TDR for RT inhibitors was identified in five out of 30 samples (16.7%). Conclusion: these results indicate the emergence of TDR among ART-naive individuals in Buleleng, Bali. This issue warrants serious consideration because TDR may hamper treatment success and reduce ART efficacy among newly diagnosed individuals. Continuous surveillance with a larger sample size is necessary to monitor TDR among ART-naive individuals.

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Latar belakang: peningkatan cakupan terapi antiretroviral (ART) merupakan penyebab utama penurunan mortalitas akibat faktor terkait acquired immune deficiency syndrome (AIDS). Akan tetapi, timbulnya transmitted drug resistance (TDR) mempersempit pilihan ART yang efektif bagi individu yang belum pernah memperoleh terapi (ART-naive), yang mana dapat menghambat kesuksesan terapi. Bali menempati peringkat ke-enam sebagai provinsi di Indonesia dengan jumlah kasus kumulatif infeksi HIV tertinggi. Tujuan penelitian ini untuk mengidentifikasi adanya TDR pada individu ART-naive di Kabupaten Buleleng, Bali.

Metode: tiga puluh sembilan individu ART-naive berpartisipasi dalam studi ini. Analisis genotipik dilakukan terhadap sampel darah yang diperoleh dari partisipan.

Hasil: 28 gen protease (PR) dan 30 gen reverse transcriptase (RT) dari 37 sampel berhasil dianalisis. Subtyping menunjukkan CRF01_AE sebagai circulating recombinant yang paling dominan di Kabupaten Buleleng, Bali. TDR terhadap inhibitor PR tidak ditemukan, akan tetapi TDR terhadap inhibitor RT teridentifikasi pada lima dari 30 individu (16,7%).

Kesimpulan: adanya TDR pada individu ART-naive di Kabupaten Buleleng, Bali, perlu diperhatikan karena dapat menghambat keberhasilan terapi dan mempersempit pilihan terapi yang efektif. Surveilans berkesinambungan perlu dilakukan untuk monitoring TDR pada individu ART-naive."

"Latar belakang: Metode konvensional untuk mengkonfirmasi infeksi HIV ialah Western blot. Namun, western blot memiliki keterbatasan yaitu kontaminasi dengan antigen selular manusia dan masalah perbedaan genetik di antara subtipe HIV-1 yang menyebabkan hasil indeterminate dan ketidakakuratan diagnosis infeksi HIV-1 subtipe CRF01_AE yang dominan di Indonesia. Pemeriksaan western blot yang tersedia di Indonesia ialah untuk diagnosis infeksi HIV-1/2 dan tidak bersifat spesifik strain. Metodologi: Pada penelitian ini digunakan protein p24 rekombinan sebagai antigen pada western blot. Dilakukan optimasi ekspresi protein p24 rekombinan HIV-1 CRF01_AE pada Escherichia coli BL21CP dan purifikasi serta western blot untuk mendapatkan informasi awal mengenai reaktivitasnya terhadap serum ODHA di Indonesia. Optimasi ekspresi dilakukan terhadap lama waktu induksi, konsentrasi IPTG, dan suhu induksi. Purifikasi dilakukan dengan metode immobilized metal-affinity chromatography (IMAC) dan sistem purifikasi Ni-NTA [Qiagen] pada kondisi native dengan optimasi pada konsentrasi imidazole dalam wash buffer. Hasil: Konfirmasi protein rekombinan dengan western blot menunjukkan bahwa ekspresi dan purifikasi protein p24 rekombinan telah optimal dan reaktif terhadap serum pasien HIV-1 positif di Indonesia. Kesimpulan: Protein p24 rekombinan dari penelitian ini dapat dikembangkan untuk uji diagnostik western blot berdasarkan subtipe CRF01_AE yang dominan di Indonesia.

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Background: Conventional method for confirmation of HIV infection is western blot. However, western blot has limitation of contamination by human cellular antigen and genetic diversity matter among the HIV-1 subtypes that showed indeterminate result and inaccuracy for the diagnosis of HIV-1 subtype CRF01_AE infection predominantly in Indonesia. The western blot available in Indonesia is for diagnosis of HIV-1/2 which is not strain spesific. This research performed the p24 recombinant protein as the antigen in western blot.

Methods: We conducted the optimization in expression of p24 recombinant protein of HIV-1 subtype CRF01_AE in Escherichia coli BL21CP and purification and the confirmation by the western blot to obtain initial information about the reactivity of this recombinant protein with ODHA (people with HIV/AIDS) in Indonesia. Expression optimization administered in the induction time, IPTG consentration used, dan the induction temperature. The purification of the p24 recombinant protein carried with the immobilized metal-affinity chromatography (IMAC) method in Ni-NTA purification system [Qiagen] in native condition with optimization in the imidazole concentration used in the wash buffer.

Result: The confirmation of recombinant protein by western blot showed the expression and purification of p24 recombinant protein has been optimized well and reactive with the Indonesian HIV-1 positive serum patient.

Conclusion: This result indicated the p24 recombinant protein can be applied for the diagnostic assay development based on predominant HIV-1 subtype CRF01_AE in Indonesia."