scholarly journals DETEKSI VIRUS AVIAN INFLUENZA SUBTIPE H5N1 DI BEBERAPA PASAR UNGGAS HIDUP DALAM WILAYAH PROVINSI JAWA BARAT SEKITARNYA

2015 ◽  
Vol 9 (1) ◽  
Author(s):  
Atik Ratnawati ◽  
NLP Indi Dharmayanti
Keyword(s):  
Polymerase Chain Reaction ◽  
Avian Influenza ◽  
Reverse Transcriptase ◽  
Ribonucleic Acid ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

Pada penelitian ini dilakukan identifikasi virus avian influenza (AI) subtipe H5N1 pada unggas dan lingkungan pasar untuk mengetahui peran pasar sebagai sumber penularan virus. Metode yang digunakan dalam penelitian ini adalah pengambilan sampel swab kloaka unggas dan lingkungan di beberapa pasar di wilayah Jawa Barat dan Tangerang. Sampel selanjutnya dilakukan isolasi ribonucleic acid (RNA) dan dilakukan reverse transcriptase polymerase chain reaction (RT-PCR) dengan menggunakan primer AI subtipe H5N1. Hasil penelitian menunjukkan bahwa virus AI/H5N1 terdeteksi pada unggas dan lingkungan pasar. Disimpulkan bahwa pasar dapat menjadi sumber penularan virus AI subtipe H5N1 terhadap unggas lainnya.

scholarly journals IDENTIFICATION OF DENGUE VIRUS SEROTYPES AT THE DR. SOETOMO HOSPITAL SURABAYA IN 2016 AND ITS CORRELATION WITH NS1 ANTIGEN DETECTION

2018 ◽  
Vol 23 (2) ◽  
pp. 151
Author(s):  
Jeine Stela Akualing ◽  
Aryati Aryati ◽  
Puspa Wardhani ◽  
Usman Hadi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Ribonucleic Acid ◽  
Rna Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Ns1 Antigen ◽  
Dengue Virus Serotypes ◽  
Polymerase Chain

Serotipe virus dengue yang beredar terus mengalami perubahan dan berbeda di setiap daerah. Pergeseran serotipe maupun genotipedi dalamnya, mempengaruhi terjadinya wabah dengue di berbagai negara. Perbedaan serotipe diduga bernasab dengan deteksi antigen(Ag) non-structural 1 (NS1), namun belum banyak penelitian yang mendukung hal tersebut. Penelitian potong lintang dikerjakan sejakFebruari-Agustus 2016 dan didapatkan 60 subjek infeksi virus dengue (IVD) dan 25 non-IVD. Ribonucleic acid (RNA) virus denguediperiksa di semua subjek menggunakan Simplexa Dengue Real-Time Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)termasuk identifikasi serotipe virus dengue dan pemeriksaan NS1 menggunakan uji cepat NS1 Panbio. Perbedaan perbandingan variabelkategorikal dianalisis dengan uji Fisher Exact. Kenasaban antara serotipe dengan deteksi Ag NS1 dianalisis dengan Chi-Kuadrat. RNAvirus dengue terdeteksi di 43 dari 60 subjek IVD (71,7%). Serotipe terbanyak adalah DENV-3 (62,8%). Pergeseran dominasi serotipetelah terjadi di Surabaya, sebelumnya dari DENV-2 ke DENV-1 dan sekarang DENV-3, kemungkinan akibat mobilitas pejamu, transporvirus dan faktor geografis. Kepekaan uji cepat NS1 75% dan kekhasan 100%. Persentase deteksi NS1 antar serotipe berbeda bermakna(p=0,002). Deteksi NS1 lebih rendah pada DENV-1 dibandingkan DENV-2 (p=0,007) ataupun DENV-3 (p=0,003). Serotipe virusdengue bernasab dengan deteksi NS1 (p=0,005). Ciri serotipe maupun genotipe virus dengue kemungkinan mempengaruhi sekresiNS1. Telah terjadi pergeseran serotipe virus dengue di pasien IVD di Surabaya sehingga diperlukan surveillance berkesinambunganuntuk memperkirakan terjadinya wabah. Serotipe bernasab dengan deteksi NS1. Salah satu penyebab hasil negatif palsu NS1 adalahperbedaan serotipe.

scholarly journals PENENTUAN SUBTIPE VIRUS AVIAN INFLUENZA DENGAN METODE SINGLE STEP MULTIPLEX REVERSE TRANSCRIPTASE- POLYMERASE CHAIN REACTION (RT-PCR) ISOLAT ASAL PROVINSI ACEH

2014 ◽  
Vol 8 (1) ◽  
Author(s):  
Teuku Zahrial Helmi ◽  
Rini Widayanti ◽  
Aris Haryanto
Keyword(s):  
Polymerase Chain Reaction ◽  
Avian Influenza ◽  
Reverse Transcriptase ◽  
Influenza A ◽  
Deoxyribonucleic Acid ◽  
Single Step ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Virus Influenza

Tujuan dari penelitian ini adalah mengidentifikasi keberadaan gen M, H5, dan N1 virus avian influenza (AI) melalui metode single step multiplex reverse transcriptase-polymerase chain reaction (RT-PCR), sebagai acuan untuk peneguhan diagnosis secara molekuler virus AI di Provinsi Aceh. Penelitian ini menggunakan 11 isolat virus AI asal Provinsi Aceh yang diperoleh dari Laboratorium Balai Penyidikan dan Pengujian Veteriner (BPPV) Regional I Medan di Sumatera Utara dari tahun 2006-2008. Penelitian dilakukan di Laboratorium Virologi BPPV Regional I di Medan, Sumatera Utara. Amplifikasi terhadap gen matriks (M) virus AI menggunakan metode simplex RT-PCR. Hasil simplex RT-PCR terhadap gen M diperoleh 10 isolat yang menunjukkan pita deoxyribonucleic acid (DNA) pada 276 bp dan satu isolat yang tidak muncul, kemudian dilanjutkan dengan metode single step multiplex RT-PCR menggunakan pasangan primer gen penyandi protein N1, H5, dan M. Produk PCR 131 bp (N1), 189 bp (H5), dan 276 bp (M) muncul sebagai hasil elektroforesis dari semua isolat virus AI. Semua virus AI yang mewabah dari tahun 2006-2008 di Provinsi Aceh termasuk ke dalam virus influenza A subtipe H5N1.

scholarly journals A meta-analysis of accuracy and sensitivity of chest CT and RT-PCR in COVID-19 diagnosis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fatemeh Khatami ◽  
Mohammad Saatchi ◽  
Seyed Saeed Tamehri Zadeh ◽  
Zahra Sadat Aghamir ◽  
Alireza Namazi Shabestari ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Ct Scan ◽  
Predictive Value ◽  
Meta Analysis ◽  
Chest Ct ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Chest Ct Scan

AbstractNowadays there is an ongoing acute respiratory outbreak caused by the novel highly contagious coronavirus (COVID-19). The diagnostic protocol is based on quantitative reverse-transcription polymerase chain reaction (RT-PCR) and chests CT scan, with uncertain accuracy. This meta-analysis study determines the diagnostic value of an initial chest CT scan in patients with COVID-19 infection in comparison with RT-PCR. Three main databases; PubMed (MEDLINE), Scopus, and EMBASE were systematically searched for all published literature from January 1st, 2019, to the 21st May 2020 with the keywords "COVID19 virus", "2019 novel coronavirus", "Wuhan coronavirus", "2019-nCoV", "X-Ray Computed Tomography", "Polymerase Chain Reaction", "Reverse Transcriptase PCR", and "PCR Reverse Transcriptase". All relevant case-series, cross-sectional, and cohort studies were selected. Data extraction and analysis were performed using STATA v.14.0SE (College Station, TX, USA) and RevMan 5. Among 1022 articles, 60 studies were eligible for totalizing 5744 patients. The overall sensitivity, specificity, positive predictive value, and negative predictive value of chest CT scan compared to RT-PCR were 87% (95% CI 85–90%), 46% (95% CI 29–63%), 69% (95% CI 56–72%), and 89% (95% CI 82–96%), respectively. It is important to rely on the repeated RT-PCR three times to give 99% accuracy, especially in negative samples. Regarding the overall diagnostic sensitivity of 87% for chest CT, the RT-PCR testing is essential and should be repeated to escape misdiagnosis.

scholarly journals Use of immunodot blot and multiplex reverse transcriptase-polymerase chain reaction in dengue virus detection in macerates of Aedes aegypti larvae

2012 ◽  
Vol 3 (1) ◽  
pp. 13
Author(s):  
Aline T.A. Chagas ◽  
Michelle D. Oliveira ◽  
Jose M.S. Mezencio ◽  
Eduardo A.M. Silva ◽  
Leandro L. Oliveira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Epidemiological Surveillance ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Denv Serotypes ◽  
Polymerase Chain ◽  
Pcr Techniques

The <em>Dengue virus</em> is the main arbovirus that affects man in terms of morbidity and mortality. The detection of the virus is very important for epidemiological surveillance, so here we propose to standardize and compare the immunodot blot (IDB) and multiplex reverse transcriptase-polymerase chain reaction (M-RT-PCR) techniques to detect and characterize the dengue virus (DENV) serotypes in samples of <em>Aedes aegypti</em> larvae. Thus, the IDB and M-RT-PCR techniques were standardized using macerated samples of larvae collected in nature. The use of monoclonal antibodies in IDB has not shown great results, but DENV detection through this method was possible using polyclonal antibodies. The distinction of serotypes 1, 2 and 3 was carried out by M-RT-PCR.

scholarly journals Deteksi Molekuler Virus Dengue dan Chikungunya pada Nyamuk Aedes spp. di Kecamatan Cilongok

2020 ◽  
Vol 2 (2) ◽  
pp. 181
Author(s):  
Dwi Iva Fitriana ◽  
Endang Srimurni Kusmintarsih ◽  
Trisnowati Budi Ambarningrum
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

DBD dan chikungunya merupakan salah satu penyakit yang masih menjadi masalah di Indonesia. Kecamatan Cilongok merupakan salah kecamatan endemis DBD dan pernah mengalami KLB chikungunya. Deteksi virus pada nyamuk sebelum menginfeksi manusia penting sebagai peringatan dini dalam upaya pencegahan wabah di daerah endemis. Tujuan penelitian ini adalah mengetahui infeksi virus Dengue dan Chikungunya pada nyamuk Aedes spp. yang ditangkap. Penelitian ini dilakukan di empat desa di Kecamatan Cilongok yang meliputi Desa Cilongok, Pernasidi, Kalisari, dan Jatisaba, pengambilan sampel dilakukan secara purposive. Deteksi virus Dengue dan Chikungunya pada nyamuk dilakukan menggunakan metode Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR). Hasil positifitas virus dianalisis secara deskriptif untuk menggambarkan potensi transmisi virus. Hasil penelitian menunjukkan bahwa nyamuk Aedes spp. yang tertangkap tidak mengandung virus Dengue dan Chikungunya.  

scholarly journals Evaluation of a 384–Well Format for High-Throughput Real-Time Reverse Transcription Polymerase Chain Reaction for Avian Influenza Testing

2009 ◽  
Vol 21 (5) ◽  
pp. 679-683 ◽  
Author(s):  
Pamela J. Ferro ◽  
Jason Osterstock ◽  
Bo Norby ◽  
Geoffrey T. Fosgate ◽  
Blanca Lupiani
Keyword(s):  
Polymerase Chain Reaction ◽  
Avian Influenza ◽  
Real Time ◽  
High Throughput ◽  
Reverse Transcription ◽  
Virus Isolation ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Response Planning ◽  
Polymerase Chain

As concerns over the global spread of highly pathogenic avian influenza H5N1 have heightened, more countries are faced with increased surveillance efforts and incident response planning for handling a potential outbreak. The incorporation of molecular techniques in most diagnostic laboratories has enabled fast and efficient testing of many agents of concern, including avian influenza. However, the need for high-throughput testing remains. In this study, the use of a 384–well format for high-throughput real-time reverse transcription polymerase chain reaction (real-time RT-PCR) testing for avian influenza is described. The analytical sensitivity of a real-time RT-PCR assay for avian influenza virus matrix gene with the use of both 96– and 384–well assay formats and serial dilutions of transcribed control RNA were comparable, resulting in similar limits of detection. Of 28 hunter-collected cloacal swabs that were positive by virus isolation, 26 (92.9%) and 27 (96.4%) were positive in the 96– and 384–well assays, respectively; of the 340 hunter-collected swabs that were negative by virus isolation, 45 (13.2%) and 23 (6.8%) were positive in the 96– and 384–well assays, respectively. The data presented herein supports the utility of the 384–well format in the event of an avian influenza outbreak for high-throughput real-time RT-PCR testing.

scholarly journals A reverse transcriptase-polymerase chain reaction detects heterogeneous chimeric mRNAs in leukemias with 11q23 abnormalities

Blood ◽  
1994 ◽  
Vol 83 (10) ◽  
pp. 2912-2921 ◽  
Author(s):  
K Yamamoto ◽  
M Seto ◽  
S Iida ◽  
H Komatsu ◽  
N Kamada ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Response To Therapy ◽  
Chromosome 9 ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Single Clone ◽  
Sensitivity Studies ◽  
Polymerase Chain ◽  
11Q23 Abnormalities

Abstract The MLL gene involved in 11q23 translocations found in the majority of infantile leukemias and some secondary leukemias makes fusion transcripts with genes such as LTG4 (chromosome 4), LTG9 (chromosome 9), and LTG19 (chromosome 19) as a result of reciprocal translocation. We have examined 25 cases of leukemias with 11q23 abnormalities by Southern blot analysis and the reverse transcriptase-polymerase chain reaction (RT-PCR). Using various primer pairs, chimeric mRNAs could be amplified in 6 of 7 leukemias with t(4;11), 6 of 8 leukemias with t(9;11) including secondary leukemia, 8 of 9 leukemias with t(11;19), and 1 with a deletion at 11q23. The chimeric mRNAs were heterogeneous and differential usage of the MLL exons was found, irrespective of the partner chromosomes. Sensitivity studies showed that a single clone with chimeric mRNA in 10(4) to 10(5) cells could be detected. These findings show that the present RT-PCR settings provide a rapid, accurate, and sensitive tool for diagnosing leukemias with 11q23 translocations and for monitoring response to therapy in these patients.

Detection of Occult Melanoma Cells in Paraffin-Embedded Histologically Negative Sentinel Lymph Nodes Using a Reverse Transcriptase Polymerase Chain Reaction Assay

2001 ◽  
Vol 19 (5) ◽  
pp. 1437-1443 ◽  
Author(s):  
Giuseppe Palmieri ◽  
Paolo A. Ascierto ◽  
Antonio Cossu ◽  
Nicola Mozzillo ◽  
Maria L. Motti ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Prognostic Significance ◽  
Primary Melanoma ◽  
Disease Stage ◽  
Occult Metastasis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain

PURPOSE: Detection of occult metastasis before the development of clinical disease could allow more accurate staging, appropriate follow-up procedures, and adjuvant therapies in patients with malignant melanoma (MM). The sentinel lymph node (SLN) has been proposed as a reliable predictor of metastatic disease in the lymphatic basin draining the primary melanoma. In this study, we screened both paraffin-embedded SLNs and peripheral-blood (PB) samples from MM patients at various stage of disease using a multimarker reverse transcriptase polymerase chain reaction (RT-PCR) assay. The prognostic significance of the presence of PCR-positive markers was also evaluated. PATIENTS AND METHODS: Total RNA was obtained from paraffin-embedded SLN sections and PB samples of 75 MM patients. RT-PCR was performed using tyrosinase and MelanA/MART1 as melanoma-associated markers. Radiolabeled PCR products were analyzed on denaturing polyacrylamide gels. RESULTS: Good sensitivity of the RT-PCR assay on archival tissues was demonstrated after comparison of RT-PCR results on frozen and paraffin-embedded SLNs from 16 MM patients. Significant correlation between the disease stage and marker expression in both PB and SLN samples was observed; the highest value was for patients who were positive for both markers in SLN (P = .006). Progression of disease was significantly associated with the total number of PCR-positive markers in both PB (P = .034) and SLN (P = .001) samples. CONCLUSION: Although sensitivity is lowered by the use of paraffin-embedded specimens, our data indicate that RT-PCR analysis of serial sections from archival SLNs may be helpful in improving detection of occult micrometastases, thus improving staging of patients with melanoma.

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