scholarly journals Assessment of Developmental and Reproductive Fitness of Dengue-Resistant Transgenic Aedes aegypti and Improvement of Fitness Using Antibiotics

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Hewawasam Patuwatha Badathuruge Kalindu Dulanja Ramyasoma ◽  
Yasanthi Illika Nilmini Silva Gunawardene ◽  
Menaka Hapugoda ◽  
Ranil Samantha Dassanayake
Keyword(s):  
Dengue Virus ◽  
Virus Transmission ◽  
Reproductive Fitness ◽  
Animal Populations ◽  
Virus Rna ◽  
Virus Serotype ◽  
Fitness Parameters ◽  
Level 2 ◽  
Transgenic Mosquitoes ◽  
Transgenic Mosquito

Background. Genetic modification offers opportunities to introduce artificially created molecular defence mechanisms to vector mosquitoes to counter diseases causing pathogens such as the dengue virus, malaria parasite, and Zika virus. RNA interference is such a molecular defence mechanism that could be used for this purpose to block the transmission of pathogens among human and animal populations. In our previous study, we engineered a dengue-resistant transgenic Ae. aegypti using RNAi to turn off the expression of dengue virus serotype genomes to reduce virus transmission, requiring assessment of the fitness of this mosquito with respect to its wild counterpart in the laboratory and semifield conditions. Method. Developmental and reproductive fitness parameters of TM and WM have assessed under the Arthropod Containment Level 2 conditions, and the antibiotic treatment assays were conducted using co-trimoxazole, amoxicillin, and doxycycline to assess the developmental and reproductive fitness parameters. Results. A significant reduction of developmental and reproductive fitness parameters was observed in transgenic mosquito compared to wild mosquitoes. However, it was seen in laboratory-scale studies that the fitness of this mosquito has improved significantly in the presence of antibiotics such as co-trimoxazole, amoxicillin, and doxycycline in their feed. Conclusion. Our data indicate that the transgenic mosquito produced had a reduction of the fitness parameters and it may lead to a subsequent reduction of transgenic vector density over the generations in field applications. However, antibiotics of co-trimoxazole, amoxicillin, and doxycycline have shown the improvement of fitness parameters indicating the usefulness in field release of transgenic mosquitoes.

scholarly journals Detection of Dengue Virus RNA in Patients after Primary or Secondary Dengue Infection by Using the TaqMan Automated Amplification System

1999 ◽  
Vol 37 (8) ◽  
pp. 2543-2547 ◽  
Author(s):  
Thomas Laue ◽  
Petra Emmerich ◽  
Herbert Schmitz
Keyword(s):  
Viral Load ◽  
Dengue Virus ◽  
Dengue Infection ◽  
Immunoglobulin M ◽  
Viral Rna ◽  
Serum Samples ◽  
Igg Antibody ◽  
Igm Antibodies ◽  
Virus Rna ◽  
Virus Serotype

In consecutive serum samples from 25 tourists with acute dengue fever, virus-specific RNA was detected by using fully automated TaqMan reverse transcriptase PCR. For this amplification technique new primers and special fluorochrome-labeled probes had to be synthesized. During amplification the increasing amount of viral DNA could simultaneously be measured in the tightly sealed tubes. Dengue virus RNA was found in almost all patients (17 of 18), if the samples had been taken soon after the onset of symptoms and before anti-dengue virus antibody had been produced. RNA was detectable in only one of five persons who had anti-dengue virus immunoglobulin M (IgM) antibodies but not yet IgG antibodies. In 30 late samples with both IgG and IgM antibodies viral RNA was no longer demonstrable. In two early samples from two frequent travelers obtained 1 and 2 days after the onset of symptoms significant IgG antibody titers were present but there were no anti-dengue virus IgM antibodies. In these samples a viral load of >5 × 106 dengue virus RNA copies (dengue types 1 and 2) was detectable. These findings of a high viral load in the presence of anti-dengue virus IgG antibody are suggestive of a secondary dengue virus infection. In the 20 tourists (17 plus 1 plus 2) in whom viral RNA was found, the dengue virus serotype could be related to the area where the infection had taken place. Most of our patients came from southeast Asia and most frequently had dengue virus type 1 infections (8 of 20).

scholarly journals Frequency of Dengue Virus Serotype 1 in Lahore by In-house assay

2020 ◽  
pp. 1-7
Author(s):  
Hajra Farooq ◽  
Waheed Uz Zaman Tariq ◽  
Sadia Bano ◽  
Ali Raza ◽  
Omar Rasheed Chughtai ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Dengue Virus ◽  
Real Time Pcr ◽  
Fast Track ◽  
Virus Rna ◽  
Virus Serotype ◽  
Serotype 1 ◽  
Dengue Virus Serotype 1 ◽  
Commercial Kit ◽  
Dengue Epidemic

Abstract Dengue is an important systemic viral infection that is caused by the dengue virus. Ribonucleic acid (RNA) from dengue NS1 positive samples, collected randomly during dengue epidemic from October 2016 to October 2017 at Chugtai Lab, was extracted for nucleic acid. Both the detection and serotyping of dengue samples were performed using real-time PCR on Rotor Gene Q. From the 70 NS1 positive samples, 57 (81.4%) samples were confirmed to be positive for dengue virus RNA, while the remaining 13 (18.6%) were negative. Serotype 1 (DEN-1) were verified among all samples by in-house assay and using commercial kit FTD (Fast Track Diagnostics) dengue differentiation; it was concluded that our in-house assay is in 100% concordance with commercial kit. Serotype 2 (DEN-2) and serotype 3 (DEN-3) have been documented in Pakistan since 1994. Continuous...  

scholarly journals RANCANGAN PRIMER UNTUK DETEKSI VIRUS DENGUE SEROTIPE DENV-3 DAN DENV-4 DENGAN METODE NASBA DAN LFIA

2021 ◽  
Vol 13 (1) ◽  
pp. 1-10
Author(s):  
Dhian Prastowo ◽  
Asmarani Kusumawati ◽  
Triwibowo Ambar Garjito ◽  
Sitti Rahmah Umniyati ◽  
Mega Tyas Prihatin
Keyword(s):  
Nucleic Acid ◽  
Dengue Virus ◽  
Rna Virus ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nucleic Acid Sequence ◽  
Probe Design ◽  
Virus Rna ◽  
Virus Serotype ◽  
Dengue Virus Serotypes

Simple, fast, and accurate early detection is expected to reduce the mortality rate due to dengue infection. The dengue virus RNA detection method using Nucleic Acid Sequence-Based Amplification (NASBA) is an alternative method that can reduce the use of a thermocycler. The detection of NASBA amplicons was carried out using the Lateral Flow Assay (LFIA). This study was conducted to prove the effectiveness of the new primer design to detect dengue virus serotypes DENV-3 and DENV-4. In addition, this study was also conducted to measure the sensitivity and specificity of the LFIA method to detect dengue virus serotypes DENV-3 and DENV-4. The initial stage of this research was the isolation of dengue virus RNA from C6/36 cell cultures, then proceeded to design primers for NASBA and LFIA probes. NASBA reactions were performed on dengue virus serotypes DENV-3 and DENV-4 and DENV-4. The NASBA reaction products were then visualized on LFIA and agarose gel electrophoresis. The NASBA method with a new primary design can be used for the detection of dengue virus serotypes DENV-3 and DENV-4 as evidenced by electrophoresis results bands at 196 bp and 144 bp. The LFIA method with probe design can be used for the detection of dengue virus serotype DENV-3 and DENV-4 dengue virus serotype with a positive line on the test line on LFIA. The LFIA method for the detection of the dengue virus has a sensitivity up to a concentration of 10-3 (1 g/μl). The results of this study indicate that the newly designed primers are specific and sensitive for DENV-3 and DENV-4 dengue virus serotypes detection. Abstrak Deteksi dini yang sederhana, cepat dan akurat diharapkan dapat mengurangi tingkat kematian akibat infeksi dengue. Metode deteksi RNA virus dengue dengan Nucleic Acid Sequence-Based Amplification (NASBA) merupakan metode alternatif yang dapat mengurangi penggunaan thermocycler. Deteksi amplikon hasil NASBA dilakukan dengan Lateral Flow Assay (LFIA). Studi ini dilakukan untuk membuktikan efektivitas desain baru primer untuk mendeteksi virus dengue serotipe DENV-3 dan DENV-4. Di samping itu, studi ini juga dilakukan untuk mengukur sensitivitas dan spesifisitas metode LFIA untuk mendeteksi virus dengue serotype DENV-3 dan DENV-4. Tahap awal penelitian ini adalah isolasi RNA virus dengue dari kultur sel C6/36, kemudian dilanjutkan dengan merancang primer untuk NASBA dan probe LFIA. Reaksi NASBA dilakukan pada virus dengue serotipe DENV-3 dan DENV-4 dan DENV-4. Produk reaksi NASBA kemudian divisualisasikan pada LFIA dan elektroforesis gel agarosa. Metode NASBA dengan desain primer baru dapat digunakan untuk deteksi virus dengue serotipe DENV-3 dan DENV-4 yang dibuktikan oleh pita  hasil elektroforesis pada 196 bp dan 144 bp. Metode LFIA dengan desain probe dapat digunakan untuk deteksi virus dengue serotipe DENV-3 dan DENV-4 serotipe virus dengue dengan garis positif pada garis uji pada LFIA. Metode LFIA untuk deteksi virus dengue memiliki sensitivitas hingga konsentrasi 10-3 (1 μg/μl). Hasil penelitian ini menunjukkan bahwa primer baru yang dirancang bersifat spesifik dan sensitif untuk deteksi serotipe virus dengue DENV-3 dan DENV-4.

scholarly journals ANALISIS GENETIK GEN NONSTRUKTURAL 3 DENGUE VIRUS SEROTYPE 4 STRAIN INDONESIA

2019 ◽  
pp. 13-24
Author(s):  
Linlin Haeni ◽  
Beti Ernawati Dewi
Keyword(s):  
Dengue Virus ◽  
C Protein ◽  
Virus Rna ◽  
Virus Serotype

Demam Berdarah Dengue (DBD) adalah penyakit yang disebabkan virus dengan vektor nyamuk yang paling cepat menyebar di dunia. Penyebab DBD adalah virus RNA famili flaviviridae yang disebut virus dengue (DENV). Genom DENV terdiri dari tiga protein struktural yaitu capsid (C), protein membran (prM), dan protein envelop (E) serta tujuh gen protein nonstuktural yaitu NS1, NS2a, NS2b, NS3, NS4a, NS4b, dan NS5. Protein NS3 mengandung epitop yang dapat dikenali oleh sistem imun humoral maupun selular oleh karena itu protein NS3 merupakan target potensial bagi pengembangan vaksin dengue. Penelitian ini diawali dengan sekuensing pada gen NS3 DENV-4 IDS 96/10. Dari hasil sekuensing dilakukan analisis filogenetik dan analisis epitop. Analisis filogenetik menunjukkan gen NS3 IDS 96 /10 berada dalam satu clade dengan strain yang diisolasi dari Cina (2010), Singapura (2010) dan Thailand (2000). Pada gen NS3 DENV-4 IDS 96/10 terdapat epitop yang dapat dikenali oleh sel limfosit T CD4+ yaitu epitop #3 pada posisi asam amino (213-227) , #9A(243-257), #4(251-265), #5(258-272), # 6(266-280), #7(273-287) yang mempunyai urutan asam amino sama antar strain yang dibandingkan. Pada posisi epitop #8(281-295) terdapat variasi urutan asam amino. Asam amino pada posisi 500-508 dikenali oleh sel limfosit T CD8+ mempunyai urutan yang sama antar strain yang dibandingkan, dan asam amino pada posisi 526-531yang dikenali oleh limfosit B mempunyai urutan asam amino yang sama antar strain yang dibandingkan. Pengenalan epitop-epitop tersebut oleh limfosit T dan limfosit B menjadi dasar pengembangan vaksin khususnya vaksin yang khusus untuk strain Indonesia

scholarly journals Dengue Virus Transmission from Donor to Recipient during Haploidentical Stem Cell Transplantation

IDCases ◽  
2021 ◽  
pp. e01220
Author(s):  
Anjali Yadav ◽  
Neha Rastogi ◽  
K. Upasana ◽  
Sunisha Arora ◽  
Dhwanee Thakkar ◽  
...  
Keyword(s):  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Dengue Virus ◽  
Cell Transplantation ◽  
Virus Transmission ◽  
Haploidentical Stem Cell Transplantation

scholarly journals Micafungin Inhibits Dengue Virus Infection through the Disruption of Virus Binding, Entry, and Stability

2021 ◽  
Vol 14 (4) ◽  
pp. 338
Author(s):  
Yen-Chen Chen ◽  
Jeng-Wei Lu ◽  
Chia-Tsui Yeh ◽  
Te-Yu Lin ◽  
Feng-Cheng Liu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Early Stage ◽  
Quantitative Polymerase Chain Reaction ◽  
Enterovirus 71 ◽  
Denv Infection ◽  
Immunofluorescence Assay ◽  
Dependent Manner ◽  
Virus Binding ◽  
Virus Serotype

Dengue fever is an arbovirus disease caused by infection with the dengue virus (DENV). Half of the world’s population lives under the threat of dengue fever, however, researchers have yet to develop any drugs that are clinically applicable to this infection. Micafungin is a member of the echinocandins family of anti-fungal drugs, capable of blocking the synthesis of β-1,3-D-glucan in the walls of fungal cells. Previous studies have demonstrated the effectiveness of Micafungin against infections of enterovirus 71 (EV71) and chikungunya virus (CHIKV). This is the first study demonstrating the effectiveness of micafungin in inhibiting the cytopathic effects of dengue virus serotype 2 (DENV-2) in a dose-dependent manner. Time-of-addition assays verified the inhibitory effects of micafungin in pre-treated, co-treated, and full-treatment groups. Binding and entry assays also demonstrated the effectiveness of micafungin in the early stage of DENV-2 infection. The virucidal efficacy of micafungin appears to lie in its ability to destroy the virion. Molecular docking assays revealed the binding of micafungin to the envelope protein of DENV-2, thereby revealing the mechanism by which micafungin affects the early stage of DENV infection and the stability of DENV. Two other micafungin analogs, caspofungin and anidulafungin, were also shown to have the antiviral effects on DENV-2. Finally, immunofluorescence assay (IFA) and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) confirmed the broad anti-DENV ability of micafungin against dengue virus serotypes 1, 3, and 4 (DENV-1, DENV-3, and DENV-4). Taken together, these results demonstrate the potential of micafungin and its analogs as candidates for the development of broad-spectrum treatments for DENV infection.

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