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"Measles merupakan salah satu penyakit infeksi menular dengan jumlah kasus yang masih tinggi di Indonesia. Jumlah kasus yang tinggi tersebut perlu dilakukan konfirmasi secara laboratoris sehingga dapat dilakukan tindakan pencegahan secara cepat dan tepat. Penelitian ini menggunakan spesimen urin untuk pemeriksaan laboratorium measles dengan metode kultur virus dan Reverse Transcriptase Polymerase Chain Reaction RT-PCR. Kultur virus dilakukan dengan menggunakan sel vero/hSLAM lalu dilakukan pengamatan Cytopathic effect CPE, sedangkan RT-PCR digunakan untuk mengamplifikasi fragmen gen N menggunakan primer MeV 214 dan MeV 216. Hasil sampel didapatkan sebanyak 120 spesimen urin yang berasal dari 9 provinsi berbeda di Indonesia selama periode tahun 2016. Hasil pemeriksaan menunjukkan nilai positivitas kultur virus sebesar 7, sedangkan nilai positivitas RT-PCR sebesar 36. Hasil penelitian menunjukkan bahwa metode RT-PCR memiliki nilai positivitas yang lebih tinggi dalam mendeteksi virus measles dibandingkan dengan kultur virus.

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Measles is one of infectious diseases with a high number of cases in Indonesia. The high number of cases needs to be confirmed in a laboratory so that precautions can be taken quickly and accurately. This study used urine specimens for laboratory measles examination using viral culture method and Reverse Transcriptase Polymerase Chain Reaction RT PCR. Viral culture was done by using vero cells hSLAM then made observations cytopathic effect CPE, while RT PCR were used to amplify the N gene fragment using the primers 214 MeV and MeV 216. The results showed a number of 120 specimens of urine obtained from 9 different provinces in Indonesia during the period of 2016. the test results showed a virus culture positivity value of 7 while the value of RT PCR positivity of 36. The results showed that the RT PCR method has a higher positivity value in detecting measles virus compared to viral culture."

"Penelitian deteksi fragmen gen NA virus avian influenza (AI) subtipe H5N1 telah dilakukan menggunakan 4 pasang primer NA yang dirancang oleh peneliti di Laboratorium IHVCB, yaitu NIF64 + NIR320, NIF306 + NIR537, NIF600 + NIR774, dan NIF757 + NIR975. Tujuan penelitian mengetahui spesifisitas dan sensitivitas primer yang telah dirancang dalam mendeteksi gen NA virus AI subtipe H5N1. Teknik two-step RT-PCR digunakan untuk mendeteksi gen NA virus AI subtipe H5N1. Uji spesifisitas PCR keempat pasangan primer dilakukan menggunakan sampel virus influenza A/chicken/Indonesia/2005 (H5N1); A/chicken/Indonesia/2006 (H5N1); A/chicken/Indonesia/2007 (H5N1); A/Indonesia/2007 (H3N2); A/Indonesia/2007 (H1N1); bakteri Streptococcus pneumonia, Neisseria meningitidis, dan Haemophilus influenzae. Uji sensitivitas PCR dilakukan dengan pengenceran bertahap terhadap cDNA virus influenza A/chicken/Indonesia/2006 (H5N1) dengan nilai konsentrasi 0,1 pg/μl--10 ng/μl. Hasil penelitian menunjukkan keempat pasangan primer memiliki spesifisitas tinggi terhadap virus AI subtipe H5N1, tidak pada subtipe H1N1 dan H3N2, serta tidak terjadi reaksi silang antara primer dan gen bakteri patogen saluran pernapasan. Pasangan primer NIF306 + NIR537 mempunyai sensitivitas PCR paling tinggi dibandingkan ketiga pasangan primer lainnya karena dapat mendeteksi gen NA dengan nilai konsentrasi cDNA terendah sebesar 1 pg/μl sehingga paling baik digunakan pada uji diagnostik AI dengan RT-PCR."

"Wabah infeksi virus banyak terjadi di lingkungan fasilitas kesehatan, seperti puskesmas. Transmisi virus di lingkungan puskesmas ini tidak hanya memberikan dampak buruk kepada pasien, namun juga kepada perawat maupun dokter yang bekerja di puskesmas.Tujuan dari penelitian ini adalah untuk mendeteksi serta mengetahui ada atau tidaknya asam nukleat milik Respiratory Syncytial Virus (RSV) dan Enterovirus 71 di lingkungan Puskesmas Ciracas, Jakarta Timur menggunakan Reverse Transcription- Polymerase Chain Reaction (RT-PCR). Permukaan benda pengambilan sampel dipilih berdasarkan kemungkinan sering kontak langsung dengan pengunjung Puskesmas dan kemungkinan terjadinya transmisi virus. Sampel diambil menggunakan metode swab, yang kemudian dilakukan proses ekstraksi RNA, dan sintesis cDNA dengan bantuan enzim Reverse Transcriptase. Sampel selanjutnya dapat digunakan untuk proses PCR dan elektroforesis. Total 32 sampel semua menunjukkan hasil yang negatif, yaitu tidak ditemukan asam nukleat milik RSV dan EV-71 di sampel. Hal ini dapat disebabkan oleh adanya protokol kebersihan yang ketat di Puskesmas Kecamatan Ciracas yang sudah dijalankan dengan baik untuk meminimalkan kontaminasi virus ke lingkungan Puskesmas.

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Outbreaks of virus can often occur in healthcare settings, such as primary health care. Transmission virus in primary health care environment represents a serious risk not only for patients, also for both staff and doctor. The aim of this study was to detection of Respiratory Syncytial Virus (RSV) and Enterovirus 71 nucleic acids on Environmental Surface in Ciracas Primary Health Care, East Jakarta using Reverse Transcription Polymerase Chain Reaction (RT-PCR). The following sampling sites have been recommended based on high-touch surfaces with health visitors and possible transmission virus routes. The samples was taken using swab, and then used samples for RNA extraction and cDNA synthesis by using Reverse Transcription enzyme. The samples can then be used in PCR and electophoresis. In total 32 surface samples were collected and 32 surface samples tested negative for both RSV and Enterovirus 71 nucleic acids. The negative result caused by effective hygiene procedures have been applied in Ciracas Primary Health Care to prevent and minimize the contamination and spread of the virus in environment."

"Measles immunization has been introduced since 1960, thereby markedly reducing the number of cases in developed countries. However, measles epidemics still occur even in developed countries. In the United States, in 1988-1992 an increase in the number of measles cases reaching 50,000 cases was reported. Some of these cases occurred in previously immunized patients. This was thought to be caused by genetic mutation of the measles virus, aside from weaknesses of the vaccine and low immunization coverage.Since measles immunization was employed in Indonesia, the number of measles patients has decreased. However, epidemics are still frequently reported. About 15-30% of reported cases occurred in those previously immunized, raising the question of whether a genetic difference exists between the wild-type measles virus circulating in Indonesia and the vaccine virus being used. Such a difference may lead to the differences in the antigenicity of the wild-type and vaccine viruses, rendering the resulting antibody incapable of neutralizing the wild-type viruses. Based on the above, this study is aimed to demonstrate the extent of genetic and antigenic differences between the wild-type and vaccine measles viruses.We conducted an experimental laboratory study to sequence the N, H, and F genes of the wild-type measles viruses (G2, G3, and D9) and the CAM-70 vaccine virus. To show antigenic differences, the wild-type viruses (G2, G3, and D9) and the CAM-70 and Schwarz viruses were injected to BALB/c mice. Serum antibodies of the mice were analyzed using ELISA, cross-neutralization test, and immunoblotting using antigens from the respective viruses.Results of this study showed that the wild-type and the vaccine viruses differ in the sequence of the N gene by 73-79 nucleotides, resulting in amino acid substitution of 17-24 residues; the H gene by 60-99 nucleotides, resulting in amino acid substitution of 13-29 residues; the F gene by 71-88 nucleotides, resulting in amino acid substitution of 4-3 I residues. Differences between the wild-type and the CAM-70 and Schwarz vaccine viruses were also found in the epitope site of the CTL and antibodies, which are important to virus antigenicity.We conclude that a significant difference in antigenicity exists between the wild-type measles viruses circulating in Indonesia with the CAM-70 measles virus. We also found the immunogenicity of the CAM-70 and Schwarz vaccine viruses to be lower than that of the wild-type viruses."

"Measles immunization has been introduced since 1960, thereby markedly reducing the number of cases in developed countries. However, measles epidemics still occur even in developed countries. In the United States, in 1988-1992 an increase in the number of measles cases reaching 50,000 cases was reported. Some of these cases occurred in previously immunized patients. This was thought to be caused by genetic mutation of the measles virus, aside from weaknesses of the vaccine and low immunization coverage. Since measles immunization was employed in Indonesia, the number of measles patients has decreased. However, epidemics are still frequently reported. About 15-30% of reported cases occurred in those previously immunized, raising the question of whether a genetic difference exists between the wild-type measles virus circulating in Indonesia and the vaccine virus being used. Such a difference may lead to the differences in the antigenicity of the wild-type and vaccine viruses, rendering the resulting antibody incapable of neutralizing the wild-type viruses. Based on the above, this study is aimed to demonstrate the extent of genetic and antigenic differences between the wild-type and vaccine measles viruses. We conducted an experimental laboratory study to sequence the N, H, and F genes of the wild-type measles viruses (G2, G3, and D9) and the CAM-70 vaccine virus. To show antigenic differences, the wild-type viruses (G2, G3, and D9) and the CAM-70 and Schwarz viruses were injected to BALB/c mice. Serum antibodies of the mice were analyzed using ELISA, cross-neutralization test, and immunoblotting using antigens from the respective viruses. Results of this study showed that the wild-type and the vaccine viruses differ in the sequence of the N gene by 73-79 nucleotides, resulting in amino acid substitution of 17-24 residues; the H gene by 60-99 nucleotides, resulting in amino acid substitution of 13-29 residues; the F gene by 71-88 nucleotides, resulting in amino acid substitution of 4-3 I residues. Differences between the wild-type and the CAM-70 and Schwarz vaccine viruses were also found in the epitope site of the CTL and antibodies, which are important to virus antigenicity. We conclude that a significant difference in antigenicity exists between the wild-type measles viruses circulating in Indonesia with the CAM-70 measles virus. We also found the immunogenicity of the CAM-70 and Schwarz vaccine viruses to be lower than that of the wild-type viruses."

"Campak adalah penyakit virus akut(paramyxavirus) sangat mudah menular melalui udara atau kontak langsung namun tergolong penyakit yang dapat dicegah dengan imunisasi. Di Indonesia penyakit campak telah masuk pada tahap reduksi dengan cakupan imunisasi (>90 %) namun Case fatality rate (CFR) eukup tinggi yaitu sekitar 1,7 - 2,4 oleh karena itu penelitian kearah mencari faktor penyebab penyakit campak pads balita dalam hal ini dibatasi pada faktor kesehatan lingkungan dan karakteristik anak balita yang berkaitan dengan kejadian penyakit campak pada balita menjadi sangat beralasan.Penelitian ini bertujuan untuk mengetahui distribusi frekwensi, hubungan dan mencari model faktor kesehatan lingkungan (16 variabel) dan karakteristik anak balita (5 variabel ) dengan kejadian penyakit campak pada balita. Penelitian dilaksanakan di Kabupaten Garut dengan metode kasus kontrol, jumlali sampel masing masing 150 kasus dan 150 kontrol total 300 sampel (1:1), rentang waktu antara Bolan Juli 2000 aid Bulan Desember 2001.Hasil penelitian menunjukan bahwa dari 21 Variabel yang dilakukan uji hubungan bevariat ada 15 variabel yang memiliki hubungan bermakna dengan p 0.05 (hipotesis ditolak). Dan 5 variabel p > 0.05 (hipotesis gagal ditolak).Model akhir tanpa interaksi didapat lima variabel utama yang berhubungan dengan kejadian campak adalah Imunisasi nilai B (3.340), Jendela (1.468), Vit A ( 1.319), Kepadatan ( 0.885) dan Cahaya (0.846) dengan konstanta -5.218. Faktor paling dominan adalah imunisasi dengan OR 28.228 pada CI 95 % 11.789-67.588, sedangkan setelah melalui uji interaksi terdapat dua variabel tunggal dan 2 yang berinteraksi yaitu 1286 (Imunisasi), 1,393 (Cahaya by Jendela), 0.933 (Kepadatan), dan 0.947 (Cahaya by Vit A) dengan konstanta -3.951 faktor paling dominan yang dapat mempengaruhi kejadian campak adalah Imunisasi dengan nilai B = 3.951 dengan QR = 26.72 nilai C195 % = 11.301-63.201Untuk aplikasi penanganan program ini tentu memerlukan strategi khusus, yang intinya perlu pelayanan kesehatan masyarakat yang komprehensif berupa pelayanan promosi dan pencegahan berupa pelayanan intensif pelaksanaan imunisasi dan pemberian vitamin A serta melaksanakan perbaikan kesehatan lingkungan fisik rumah terutama sistem pencahayaan, jendelanisasi, dan pengurangan kepadatan kamar.

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Measles is an accute viruses deseases (paramyxovirus)_ It is very easy infected to other people direct contact, but can be prevented by immunization. In Indonesia measles deseases is in reduction phase with immunimtion trap >90 %, but the Case fatality rate (CFR) is high between 1.7 - 2.4. There efor the study to find the risk factor of measles on childhood in this case is limited on environtmenal health factor and the characteristic of childhood that is associated with measles incidence of childhood is very reasonable.

The purpose of this study is to know the distribution anda freqkfency, the association and find the environment health factors model (16 variables) and characteristik of childhood (5 variable) with the measles incidence on Childhood at Garut District 2000-2001 year.

This study was being done at Garut district using case control method_ The sample of this study is 300 ehilldhood (150 cases and 150 control) the study last from July 2000 --- Descember 2000. The result of this study showed that from 21 variable there is 16 variabels is significant because p < 0.05.

The multivariate final model are : immunization B velue (3.340), Windows (1.468), Vit A ( 1.319), Crowding ( 0.885) and Lighting (0.846), constanta -5.218. The strenght of Factor is immunization with OR 28.228 at CI 95 % 11389-67.588.

Interaksi test result is 3.286 (Imunisation), 1.393 (Light by windows), 0.933 (Croeding, and 0.947 (Lighting by Vit A), constanta -3.951 and strenght factor is Imunisation with B value = 3.951 , OR = 26.72 Cl 95 % = 11.301-63.201

Sugestion for program Aplication cocerning measles program in Garut District is a comprehensif action, covering Promotion, prevention, Curative dan Rehabilitation. The priority program are Immunization programe, Vitamin A, and Rehabilitation of Window, sistem of Lighting Room and reduction of Ovbercrowding."

"Latar belakang. Cakupan imunisasi campak di Indonesia mencapai 80% namun prevalens campak di Indonesia masih tinggi, terutama pada anak usia 1-4 tahun. WHO merekomendasikan pemberian imunisasi campak ke-2 pada tahun kedua kehidupan. Di Indonesia diberikan pada usia 15-18 bulan dalam kombinasi vaksin MMR. Sayangnya cakupan imunisasi MMR masih rendah sehingga Depkes merekomendasikan pemberian imunisasi campak ke-2 pada usia 2 tahun untuk meningkatkan imunitas seorang anak terhadap penyakit campak.Tujuan. Penelitian ini untuk mengetahui: (1) proporsi anak usia 1-4 tahun yang telah mendapatkan imunisasi campak 1 kali yang memiliki antibodi campak mencapai kadar protektif dan rerata kadar antibodinya, (2) proporsi anak usia 1-4 tahun yang telah mendapatkan imunisasi campak ≥ 2 kali yang memiliki antibodi campak mencapai kadar protektif dan rerata kadar antibodinya, (3) hubungan antara usia saat diperiksa kadar antibodi campak, usia saat imunisasi, status gizi, kondisi kesehatan saat imunisasi campak terhadap antibodi campak, (4) hubungan antara pemberian imunisasi campak dosis ke-dua terhadap antibodi campak.Metode. Penelitian potong lintang di 6 posyandu di 5 wilayah DKI Jakarta pada bulan Juni hingga Agustus 2014. Anak yang memenuhi kriteria inklusi diperiksa kadar IgG campak. Dari hasil pemeriksaan IgG campak, kemudian ditentukan apakah mencapai kadar protektif atau tidak dan rerata kadar antibodinya. Dicari apakah terdapat hubungan antara imunisasi campak dosis ke-dua dengan kadar antibodi campak.Hasil. Dari 145 subjek penelitian, 125 subjek (86,2%) memiliki kadar antibodi campak yang mencapai kadar protektif (≥ 120 mIU/ml) dan 20 subjek (13,8%) memiliki kadar antibodi campak yang tidak mencapai kadar protektif (< 120 mIU/ml). Median kadar antibodi campak pada kelompok protektif adalah 844 mIU/ml, dengan nilai minimum 129 mIU/ml dan nilai maksimum 5000 mIU/ml. Kelompok usia 3-4 tahun memiliki kadar antibodi campak yang mencapai kadar protektif terbanyak (91,8%) dibanding kelompok usia 2-3 tahun (88,2%) dan 1-2 tahun (72,7%). Tidak didapatkan hubungan antara usia saat mendapatkan imunisasi campak dan status gizi terhadap kadar antibodi campak.Simpulan. (1) Proporsi anak usia 1-4 tahun yang mendapatkan imunisasi campak 1 kali dan memiliki antibodi campak mencapai kadar protektif sebesar 77% (54/70) dengan median kadar antibodinya adalah 733,5 mIU/ml, (2) Proporsi anak usia 1-4 tahun yang mendapatkan imunisasi campak ≥ 2 kali dan memiliki antibodi campak mencapai kadar protektif sebesar 94,6% (71/75) dengan median kadar antibodinya adalah 885 mIU/ml. (3) Pemberian imunisasi campak ≥ 2 kali meningkatkan timbulnya antibodi campak yang mencapai kadar protektif sebesar 1,227 kali dibanding pemberian imunisasi campak 1 kali.

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Background. Indonesia measles immunization coverage reach 80% but measles prevalence remains high especially in children 1-4 years old. WHO recommended second dose of measles containing vaccine at second year of age. In Indonesia, it has been done through MMR vaccine at 15-18 month. Unfortunately MMR immunization coverage still low and Ministry of Health recommended second dose of measles containing vaccine for all 2 years old children who have never been immunized with MMR vaccine at 15-18 month to increase the immunity against measles.

Objectives. This study aimed to know: (1) proportion of children 1-4 years old who has been immunized one time measles vaccine and reach protective antibody level and mean of antibody, (2) proportion of children 1-4 years old who has been immunized twice or more measles vaccine and reach protective antibody level and mean of antibody, (3) association between age, age of immunization, nutritional status, and health status when being immunized with measles antibody level, (4) association between second dose of measles vaccine with measles antibody level.

Methods. Cross-sectional study performed in 6 posyandu in 5 region of Jakarta since June until August 2014. Children who met the inclusion criteria were checked for measles IgG, identified for reaching protective level and mean of antibody. Association between second dose of measles vaccine with measles antibody level was also measured.

Results. From 145 participants, 125 (86,2%) had protective measles antibody level (≥ 120 mIU/ml) and 20 (13,8%) had not reached protective level (< 120 mIU/ml). The median measles antibody level in protective group was 844 mIU/ml, with minimum point was 129 mIU/ml and maximum point was 5000 mIU/ml. Children in 3-4 years old group had highest percentage of protective measles antibody level (91,8%) compare to children in 2-3 years old group (88,2%) and 1-2 years old group (72,7%). There were no association between age of immunization and nutritional status with measles antibody level.

Conclusion. (1) Proportion of children 1-4 years old who has been immunized one time measles immunization and reach protective measles antibody level was 77% (54/70) with the median of measles antibody level was 733,5 mIU/ml, (2) Proportion of children 1-4 years old who has been immunized twice or more measles immunization and reach protective measles antibody level was 94,6% (71/75) with the median of measles antibody level was 885 mIU/ml, (3) Twice or more measles immunization will increase protective level of measles antibody 1,227 times compare to one time measles immunization."

"Infeksi Human Immunodeficiency Virus (HIV) masih menjadi perhatian bagi masyarakat luas karena tidak dapat disembuhkan secara sempurna. Penelitian sebelumnya menyatakan bahwa virus HIV masih dapat ditemukan di jenazah sampai beberapa waktu setelah kematian, tanpa diketahui apakah masih mampu bereplikasi dan menginfeksi orang. Karena itu, penelitian ini ingin mengetahui kemampuan replikasi virus HIV di dalam sel darah putih secara in vitro dengan meniru kondisi seperti yang terjadi pada proses setelah kematian yaitu tidak terpapar oksigen. Penelitian menggunakan disain eksperimental dengan menggunakan darah 'Orang dengan HIV-AIDS' (ODHA) yang masih hidup untuk menggantikan darah jenazah ODHA terinfeksi HIV. Hal ini dikarenakan sulitnya mendapatkan sampel darah yang dapat diteliti hingga rentang waktu 48 jam dengan suhu 26-32°C seperti suhu yang lazim terjadi pada umumnya jenazah di Indonesia. Darah terinfeksi HIV tersebut diperiksa 'viral load' dan diambil sel darah putihnya. Sel darah putih tersebut dicampur kembali dengan plasma darahnya, dan ditutup minyak goreng yang sudah dipanaskan agar tidak terjadi paparan oksigen untuk mendekati kondisi postmortem. Suspensi dikultur dan supernatannya diperiksa dengan 'Reverse Transcriptase Polymerase Chain Reaction' (RT PCR) untuk melihat hasil replikasi virus HIV. Hasil penelitian ini menunjukkan, virus HIV masih bereplikasi sampai waktu 48 jam setelah paparan oksigen dihentikan. Selain itu terdapat perubahan morfologi sel darah putih yaitu efek 'cytopathic effect' (CPE) pada sel di dalam kultur yang menunjukkan adanya infeksi antar sel.

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HIV infection is still a community problem that cannot be solved perfectly. Recent studies show HIV virus can still be found in dead bodies, altough there were no evidence that indicate HIV infection from dead bodies. This research aims to elaborate HIV virus replication in leucocyte in vitro imitating dead bodies physiologic condition of oxygen deprivation.

Research is conducted using experimental design using blood samples taken from living HIV-infected persons to substitute for HIV-infected dead bodies. This subtitution held because of the difficulty to obtain blood sample from dead bodies, and studied until 48 hours postmortem on 26-32°C. HIV-infected blood was examined for viral load. The leucocyte were separated from the blood and mixed with blood plasma. This suspension stored in the tube and the upper surface added with heated cooking oil to prevent oxygen exposure. The suspension was centrifuged, the leucocyte were cultured. After cultured, the supernatan was scanned with Reverse Transcriptase Polymerase Chain Reaction (RT PCR) to detect replication of HIV.

The replication of HIV virus were detected up to 48 hours. The study also found morphologic changes of leucocyte due to cytopathic effect which showed cell-to-cell infections."

"Lesi fokal otak merupakan komplikasi neurologi pada pasien HIV yang ditandai oleh lesi desak ruang (Space Occupying Lesion) yang membutuhkan penanganan cepat dan tepat. Di beberapa negara, lesi ini dapat disebabkan oleh toksoplasma ensefalitis dan limfoma otak primer. Lesi yang disebabkan oleh toksoplasmosis dan limfoma otak primer yang disebabkan oleh Epstein Barr virus sulit untuk dibedakan menggunakan CT scan ataupun MRI. Pemeriksaan gold standar untuk membedakan keduanya yaitu dengan biopsi otak, namun hal ini merupakan tindakan invasif dan dapat menimbulkan komplikasi. Penelitian ini bertujuan untuk memperoleh uji deteksi untuk diagnosis cepat infeksi Toxoplasma gondii dan Epstein Barr virus. Desain yang dipakai pada penelitian adalah studi eksperimental laboratorium. Uji deteksi yang dikembangkan adalah dupleks real-time PCR yang dapat mendeteksi T.gondii dan EBV atau kombinasi keduanya dalam satu reaksi pada sampel pasien HIV dengan gejala klinis tersangka infeksi otak. Tahap pertama dilakukan optimasi dupleks real-time PCR meliputi suhu annealing, konsentrasi primer dan probe, uji volume elusi dan volume cetakan. Penentuan ambang batas deteksi dilakukan untuk mengukur minimal T.gondii dan EBV yang dapat dideteksi. Reaksi silang untuk mengetahui spesifisitas teknik dilakukan menggunakan bakteri dan virus sebagai berikut Staphylococcus aureus, Klebsiella pneumonia, Pseudomonas aeruginosa, Mycobacterium tuberculosis H37Rv, Candida spp, Cytomegalo virus, Herpes zoster virus, dan Varicella zoster virus. Dupleks real-time PCR yang telah optimal diaplikasi pada sampel pasien. Sampel yang digunakan adalah darah dan cairan serebrospinal dari pasien HIV dengan gejala klinis infeksi otak yang dirawat di bagian neurologi RSCM. Hasil optimasi dupleks real-time PCR diperoleh suhu annealing untuk T.gondii dan EBV 58°C, konsentrasi primer forward dan reverse untuk T.gondii dan EBV adalah 0,2 µM, konsentrasi probe T.gondii 0,4µM, konsentrasi probe EBV 0,2 µM. Deteksi ambang batas minimal DNA untuk T.gondii 5,68 copy /ml, sedangkan EBV 1,31 copy/ml. Uji yang dikembangkan pada penelitian ini termasuk uji yang sensitif dibandingkan hasil penelitian lain. Uji reaksi silang primer dan probe dupleks real-time PCR terhadap beberapa bakteri dan virus lain, menunjukkan tidak bereaksi silang dengan primer dan probe yang digunakan untuk mendeteksi T.gondii dan EBV. Hasil pemeriksaan dupleks real-time PCR pada sampel darah diperoleh 16% positif T.gondii, 40% positif Epstein Barr virus, sebanyak 16% positif Epstein Barr virus dan T.gondii dan pada sampel cairan serebrospinal diperoleh hasil 20% positif T.gondii, sebanyak 28% positif Epstein Barr virus dan 4% positif terhadap Epstein Barr Virus dan T.gondii.

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Focal brain lesion is neurology complication in HIV that marked with Space Occupying Lesion (SOL), that need rapid and effective handling. In most country, this lesion could be cause by encephalitis toxoplasma and Primary Central Nervous System Lymphoma that related to Epstein Barr virus infection that was difficult to distinguished using CT scan or MRI. Gold standard to distinguished was brain biopsy, but this examination was invasive procedure that cause complication. Therefore, we need a reliable and rapid examination to distinguished it. This study aimed to get detection for rapid diagnosis of T.gondii and EBV infection. This study was an experimental laboratory. First step was optimation of dupleks real-time PCR include annealing temperature, primer andprobe consentration, elution volume and template volume. Minimal detection of DNA to measured minimal T.gondii and EBV that could be detected. Cross reaction to know technique spesivisity using bacterial and virus Staphylococcus aureus, Klebsiella pneumonia, Pseudomonas aeruginosa, Mycobacterium tuberculosis H37Rv, Candida spp, Cytomegalo virus, Herpes zoster virus, and Varicella zoster virus. Dupleks real-time PCR has been optimally applied to patient. The sample from blood and cerebrospinal fluid of HIV patients who admitted in the neurology department of RSCM then examined to duplex real-time PCR to detect T.gondii and EBV. The optimation of duplex real-time PCR, the annealing temperature for T.gondii and EBV were 58°C, consentration of primer forward and reverse for T.gondii and EBV were 0,2 µM, consentration of probe for T.gondii was 0,4µM and EBV was 0,2µM.. Minimal DNA detection for T.gondii was 5,68 copy/ml and EBV was 1,31 copy /ml. This study was sensitive like the others. Spesivisity technique of real-time PCR, there was not cross reaction between another bacteria and virus that used as primer and probe for T.gondii and EBV. From the results of the duplex real-time PCR on blood samples, 16 % was positive T.gondii, 40% Epstein Barr virus, and 16% were positive Epstein Barr virus and T.gondii and from cerebrospinal fluid samples 20% was positive T.gondii, 28% was positive Epstein Barr virus and 4% were positive for Epstein Barr Virus and T.gondii."

"ABSTRAKKasus 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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ABSTRAKCases 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."