scholarly journals Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR

2015 ◽  
Vol 11 (2) ◽  
Author(s):  
Nastiti Wijayanti ◽  
Hera Nirwati ◽  
Tri Wibawa ◽  
Aris Haryanto ◽  
S. Sutaryo
Keyword(s):  
Dengue Virus ◽  
Nested Pcr ◽  
Genotypic Variation ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Virus Serotype ◽  
Pcr Multiplex ◽  
Pcr Products ◽  
One Step ◽  
Nested Pcr Assays

world. We have evaluated the combination of one-step RT-PCR and multiplex nested PCR assays for detectingdengue viruses from clinical samples. Twelve patients were screened for the dengue virus, using a pair of primersthat conserve for several Flavivirus. The results showed that in 12 suspect patients, 100% were positive for Flavivirusand there are some genotypic variation among them, that indicated by several RT-PCR products higher than 511 bp,the expected product for RT-PCR. Further assay was performed to clarify the presence and serotypes of dengue virususing multiplex nested PCR. Serotyping results indicated that 83,3% of samples can be confirmed for dengue virus.Among the dengue virus positive 16,7 % are dengue-2, 16.7 % are dengue-3, and the most common 50% are dengue-4,whereas dengue-1 were not found among the patients. The combination of RT-PCR and multiplex nested PCR assaycan be used for rapid analysis dengue samples in early phase which is potentially useful for clinical, epidemiologyand also evolutionary studies.Key words: Flavivirus, dengue virus, serotype, RT-PCR, multiplex nested PCR

scholarly journals One-step real-time RT-PCR assays for serotyping dengue virus in clinical samples

2015 ◽  
Vol 15 (1) ◽  
Author(s):  
Erik Alm ◽  
Gunnel Lindegren ◽  
Kerstin Ingrid Falk ◽  
Nina Lagerqvist
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Clinical Samples ◽  
Rt Pcr ◽  
One Step ◽  
Pcr Assays

Comparison of one-step RT-PCR and a nested PCR for the detection of canine distemper virus in clinical samples

2004 ◽  
Vol 82 (1-2) ◽  
pp. 83-86 ◽  
Author(s):  
YJ SHIN ◽  
KO CHO ◽  
HS CHO ◽  
SK KANG ◽  
HJ KIM ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Canine Distemper Virus ◽  
Clinical Samples ◽  
Canine Distemper ◽  
Rt Pcr ◽  
One Step

scholarly journals NS-1 antigen positive Dengue Infection and molecular characterization of Dengue Viruses in a private Medical College Hospitalin Dhaka, Bangladesh

2018 ◽  
Vol 17 (4) ◽  
pp. 669-673
Author(s):  
Mahmuda Siddiqua ◽  
Ahmed Nawsher Alam ◽  
AKM Muraduzzaman ◽  
Tahmina Shirin
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Dengue Infection ◽  
Medical Science ◽  
Cost Effective ◽  
Rt Pcr ◽  
Dengue Virus Infection ◽  
Medical College ◽  
Virus Serotype ◽  
Ns1 Antigen

Introduction: Detection of dengue virus infection as soon as possible is critical for management of dengue virus infected patients. Immuno-chromatographic (ICT) tests are easy, cost effective method for dengue virus antigen detection.The sensitivity and specificity of ICT should compare with a gold standard test like RT-PCR. Aim of this study was to compare two test methods (ICT and RT-PCR), observe dengue serotype and seasonal impact on dengue infection.Methodology & result: The patients of Ibn Sina Medical College Hospital from October 2015 to October 2017 were tested for dengue NS1 antigen by ICT method. Out of 3201 sample tested 32.39% were found positive and 89 of which were re-tested for RT-PCR for comparison. Eighty eight of 89 NS1 positive cases showed positive by RT-PCR method giving an accuracy of 98.87%. Among the RT-PCR positive cases 45 were further analyzed for serotype. DEN-1, DEN-2 or both DEN- 1 and DEN-2 were found in 21, 23 and 1cases respectively. No cases of DEN-3 or DEN-4 were detected.Conclusion: This study showed that easily available and cost effective dengue NS1 antigen detection method (ICT) is as effective as molecular test (RT-PCR). DEN-1 and DEN-2 serotype were prevalent during last few years in Bangladesh. Continuous monitoring of dengue virus serotype is important for prevention and control of sudden epidemic by other serotype. Alert to be more during post monsoon when the peak of dengue virus infection was observed.Bangladesh Journal of Medical Science Vol.17(4) 2018 p.669-673

scholarly journals Rapid and Sensitive Detection of Noroviruses by Using TaqMan-Based One-Step Reverse Transcription-PCR Assays and Application to Naturally Contaminated Shellfish Samples

2005 ◽  
Vol 71 (4) ◽  
pp. 1870-1875 ◽  
Author(s):  
Narayanan Jothikumar ◽  
James A. Lowther ◽  
Kathleen Henshilwood ◽  
David N. Lees ◽  
Vincent R. Hill ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Nested Pcr ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Specificity And Sensitivity ◽  
Routine Monitoring ◽  
The Family ◽  
One Step ◽  
Stool Specimens ◽  
Pcr Assays

ABSTRACT Noroviruses (NoV), which are members of the family Caliciviridae, are the most important cause of outbreaks of acute gastroenteritis worldwide and are commonly found in shellfish grown in polluted waters. In the present study, we developed broadly reactive one-step TaqMan reverse transcription (RT)-PCR assays for the detection of genogroup I (GI) and GII NoV in fecal samples, as well as shellfish samples. The specificity and sensitivity of all steps of the assays were systematically evaluated, and in the final format, the monoplex assays were validated by using RNA extracted from a panel of 84 stool specimens, which included NoV strains representing 19 different genotypes (7 GI, 11 GII, and 1 GIV strains). The assays were further validated with 38 shellfish cDNA extracts previously tested by nested PCR. Comparison with a recently described real-time assay showed that our assay had significantly higher sensitivity and was at least as sensitive as the nested PCR. For stool specimens, a one-step duplex TaqMan RT-PCR assay performed as well as individual genogroup-specific monoplex assays. All other enteric viruses examined were negative, and no cross-reaction between genogroups was observed. These TaqMan RT-PCR assays provide rapid (less than 90 min), sensitive, and reliable detection of NoV and should prove to be useful for routine monitoring of both clinical and shellfish samples.

scholarly journals Performance Evaluation of Commercial Dengue Diagnostic Tests for Early Detection of Dengue in Clinical Samples

2017 ◽  
Vol 2017 ◽  
pp. 1-4 ◽  
Author(s):  
Tuan Nur Akmalina Mat Jusoh ◽  
Rafidah Hanim Shueb
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Diagnostic Test ◽  
Rapid Diagnostic Test ◽  
Dengue Infection ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Kappa Agreement

The shattering rise in dengue virus infections globally has created a need for an accurate and validated rapid diagnostic test for this virus. Rapid diagnostic test (RDT) and reverse transcription-polymerase chain reaction (RT-PCR) diagnostic detection are useful tools for diagnosis of early dengue infection. We prospectively evaluated the diagnostic performance of nonstructural 1 (NS1) RDT and real-time RT-PCR diagnostic kits in 86 patient serum samples. Thirty-six samples were positive for dengue NS1 antigen while the remaining 50 were negative when tested with enzyme-linked immunosorbent assay (ELISA). Commercially available RDTs for NS1 detection, RTK ProDetect™, and SD Bioline showed high sensitivity of 94% and 89%, respectively, compared with ELISA. GenoAmp® Trioplex Real-Time RT-PCR and RealStar® Dengue RT-PCR tests presented a comparable kappa agreement with 0.722. The result obtained from GenoAmp® Real-Time RT-PCR Dengue test showed that 14 samples harbored dengue virus type 1 (DENV-1), 8 samples harbored DENV-2, 2 samples harbored DENV-3, and 1 sample harbored DENV-4. 1 sample had a double infection with DENV-1 and DENV-2. The NS1 RDTs and real-time RT-PCR tests were found to be a useful diagnostic for early and rapid diagnosis of acute dengue and an excellent surveillance tool in our battle against dengue.

Enhancing the sensitivity of Dengue virus serotype detection by RT-PCR among infected children in India

2017 ◽  
Vol 244 ◽  
pp. 46-54 ◽  
Author(s):  
Syed Fazil Ahamed ◽  
Rosario Vivek ◽  
Shalini Kotabagi ◽  
Kaustuv Nayak ◽  
Anmol Chandele ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Rt Pcr ◽  
Virus Serotype

scholarly journals Molecular characteristics of rotavirus genotypes circulating in the south of Benin, 2016-2018

2020 ◽  
Author(s):  
Jijoho Michel Agbla ◽  
Mathew D Esona ◽  
Alidéhou Jerrold Agbankpé ◽  
Annick Capo-Chichi ◽  
Rashi Gautam ◽  
...  
Keyword(s):  
World Health ◽  
Molecular Characteristics ◽  
Rt Pcr ◽  
Predominant Genotype ◽  
Pcr Multiplex ◽  
One Step ◽  
Stool Samples ◽  
Pcr Method ◽  
Health Organization ◽  
P Type

Abstract Objective: Rotavirus remains the main causative agent of gastroenteritis in young children in countries that have not yet introduced the vaccine. In Benin, rotavirus vaccine was introduced late December 2019 into the EPI. This study aims to provide pre-vaccination era rotavirus genotyping data in Benin. These data can supplement data from the surveillance system of Ministry of Health of Benin which is supported by the World Health Organization (WHO).Results: Of the 420 diarrheal stool samples, actively collected in southern Benin from July 2016 through November 2018 from children under five years old and suffering from gastroenteritis, 167 (39.8%) samples were rotavirus EIA positive. 186 (44.3%) samples contained amplifiable rotavirus RNA detected by qRT-PCR method and were genotyped using one-step RT-PCR multiplex genotyping method. G1P[8] represents the predominant genotype (32%) followed by the G2P[4] (26%), G3P[6] (16%), G12P[8] (13%) and mixed G and P types (1%). Four samples (2%) could not be assigned both G and P type specificity.

scholarly journals DETEKSI VIRUS DENGUE SEROTIPE-3 DAN PERAN SPERMATEKA DALAM PENULARAN SECARA TRANSVENEREAL PADA NYAMUK Aedes aegypti BETINA

2019 ◽  
Vol 12 (1) ◽  
pp. 130
Author(s):  
Devita Febriani Putri ◽  
Tusy Triwahyuni
Keyword(s):  
Aedes Aegypti ◽  
Nested Pcr ◽  
Rt Pcr ◽  
One Step

Penularan transvenereal berpotensi menyebarkan virus dengue melalui perilaku kawin. Pengendalian vektor Demam Berdarah Dengue (DBD) dengan strategi perilaku kawin nyamuk secara alami telah diterapkan untuk menurunkan perluasan daerah endemis DBD. Dengan dasar tersebut, pemahaman perilaku kawin nyamuk Ae. aegypti penting untuk diketahui. Penelitian ini bertujuan untuk mendeteksi virus dengue serotipe 3 (DENV-3) pada organ spermateka nyamuk Ae. aegypti betina yang telah terinfeksi DENV-3 secara transvenereal di laboratorium. Pembedahan organ spermateka pada nyamuk betina dilakukan setelah nyamuk Ae. aegypti betina kawin dengan nyamuk Ae. aegypti jantan yang positif DENV-3. Keberadaan DENV-3 pada organ spermateka nyamuk betina dilakukan dengan melakukan pengujian pooling sampel menggunakan metode One-Step RT-PCR untuk screening virus dengue (profil pita DNA spesifik 511 bp). Sampel yang hasil pengujiannya positif virus dengue, dilanjutkan dengan metode Semi-Nested PCR untuk serotyping DENV-3 (profil pita DNA spesifik 290 bp). Hasil penelitian dari 7 sampel pooling organ spermateka dari nyamuk betina positif DENV-3 hasil penularan transvenereal nyamuk jantan positif DENV-3 secara intratorakal menunjukkan tidak ada satupun sampel yang terdeteksi adanya DENV-3. Tidak ditemukan virus DENV-3 pada organ spermateka nyamuk Ae. aegypti betina yang telah terinfeksi DENV-3 secara transvenereal pada 7 sampel yang digunakan. Perlu pengujian lebih lanjut pada organ ovarium nyamuk betina untuk memastikan mekanisme terjadinya penularan transvenereal virus dengue pada Ae. aegypti dalam upaya mencari strategi baru dalam pengendalian vektor DBD

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