Detection of hantaviruses in Brazilian rodents by SYBR-Green-based real-time RT-PCR

Araújo, Jansen de; Pereira, Aluisio José; Nardi, Marcello Schiavo; Henriques, Dyana Alves; Lautenschalager, Daniele Adriana; Dutra, Lilia Mara; Ometto, Tatiana; Hurtado, Renata; Maués, Fábio Carmona de Jesus; Nava, Alessandra; Morais, Felipe Alves; Aires, Caroline C.; Favorito, Sandra E.; Durigon, Edison Luíz

doi:10.1007/s00705-011-0968-1

Cited by 12 publications

(14 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In Brazil, there are only two studies that identified hantaviruses in non-rodents reservoirs: bats ( Diphylla ecaudata and Anoura caudifer ) and marsupials ( Micoureus paraguayanus , Monodelphis iheringi and Didelphis aurita ). Sequencing the S segment of the hantavirus from both animals revealed 95% identity with the Araraquara virus, initially described in N. lasiurus rodent [238,240]. The coexistence of these animals in the same area reinforces the possibility of interspecific transmission mentioned above.…”

Section: Hantavirus Hostssupporting

confidence: 58%

Hantavirus Reservoirs: Current Status with an Emphasis on Data from Brazil

Oliveira

Guterres

Fernandes

et al. 2014

Viruses

View full text Add to dashboard Cite

Since the recognition of hantavirus as the agent responsible for haemorrhagic fever in Eurasia in the 1970s and, 20 years later, the descovery of hantavirus pulmonary syndrome in the Americas, the genus Hantavirus has been continually described throughout the World in a variety of wild animals. The diversity of wild animals infected with hantaviruses has only recently come into focus as a result of expanded wildlife studies. The known reservoirs are more than 80, belonging to 51 species of rodents, 7 bats (order Chiroptera) and 20 shrews and moles (order Soricomorpha). More than 80genetically related viruses have been classified within Hantavirus genus; 25 recognized as human pathogens responsible for a large spectrum of diseases in the Old and New World. In Brazil, where the diversity of mammals and especially rodents is considered one of the largest in the world, 9 hantavirus genotypes have been identified in 12 rodent species belonging to the genus Akodon, Calomys, Holochilus, Oligoryzomys, Oxymycterus, Necromys and Rattus. Considering the increasing number of animals that have been implicated as reservoirs of different hantaviruses, the understanding of this diversity is important for evaluating the risk of distinct hantavirus species as human pathogens.

show abstract

Section: Hantavirus Hostssupporting

confidence: 58%

Hantavirus Reservoirs: Current Status with an Emphasis on Data from Brazil

Oliveira

Guterres

Fernandes

et al. 2014

Viruses

View full text Add to dashboard Cite

show abstract

“…The concentrations of the plasmids preparation were determined by measuring the OD at 260 nm using a Nanodrop photometer (Thermo Scientific, Waltham). The plasmid copies number per microliter were determined using the formula described elsewhere [17]. All plasmids were stored at -80°C.…”

Section: Construction Of Standard Plasmidsmentioning

confidence: 99%

“…It has significantly revolutionized the detection technique for molecular diagnosis in the field of clinical microbiology, oncology, and the estimation of the interest genes expression [13][14][15][16]. Reports on HTNV detection in captured rodents based on qPCR technology have also been investigated [17].…”

Section: Introductionmentioning

confidence: 99%

Establishment of SYBR green-based qPCR assay for rapid evaluation and quantification for anti-Hantaan virus compounds in vitro and in suckling mice

Wei

Ling

et al. 2012

Virus Genes

View full text Add to dashboard Cite

Hantaan viruses cause two severe diseases lacking efficient treatment, yet no effective prophylactic vaccines are available. Continued exploration of alternative antiviral agents to treat hantavirus-related syndromes remains compulsory. The fluorescence-based quantitative real-time PCR (qPCR) has become the touchstone for target gene quantification. In the present study, standard curves for Hantaan virus (HTNV), mouse, and human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were generated by serial 10-fold dilutions of the constructed recombinant plasmid pGEM-T/HTNV, pGEM-T/mouse-GAPDH, and pGEM-T/human-GAPDH, respectively. Comparisons between the indirect immunofluorescence assay and qPCR assay in the detection of HTNV-infected Vero E6 cells showed improved detection limit and sensitivity of latter method. To characterize the inhibitory effect of several conventional antivirals (arbidol and ribavirin) and unconventional antivirals (indomethacin and curcumin) on HTNV, the levels of viral RNAs were measured for 4 days post-treatment of HTNV-infected Vero E6 cells and 18 days post-inoculation of HTNV-infected suckling mice. Our results validated that HTNV was sensitive to ribavirin and arbidol treatment, while indomethacin and curcumin may also be therapeutically effective in treating HTNV infection. As a result, the establishment and application of qPCR may be a useful tool for the evaluation of potential antivirals for Hantaan virus infection in vitro and in vivo.

show abstract

“…In Brazil, a conventional RT-PCR was able to detect Araraquara hantavirus genomes in blood and tissue samples of humans and rodents [12]. Nevertheless, molecular techniques can evolve into faster, more sensitive methods that allow for the detection and quantitation of hantaviruses in several samples.…”

Section: Introductionmentioning

confidence: 99%

Development of a One-Step SYBR Green I Real-Time RT-PCR Assay for the Detection and Quantitation of Araraquara and Rio Mamore Hantavirus

Machado

Souza

Pádua

et al. 2013

Viruses

View full text Add to dashboard Cite

Hantaviruses are members of the family Bunyaviridae and are an emerging cause of disease worldwide with high lethality in the Americas. In Brazil, the diagnosis for hantaviruses is based on immunologic techniques associated with conventional RT-PCR. A novel one-step SYBR Green real-time RT-PCR was developed for the detection and quantitation of Araraquara hantavirus (ARAV) and Rio Mamore hantavirus (RIOMV). The detection limit of assay was 10copies/µL of RNA in vitro transcribed of segment S. The specificity of assay was evaluated by melting curve analysis, which showed that the Araraquara virus amplified product generated a melt peak at 80.83 ± 0.89 °C without generating primer-dimers or non-specific products. The assay was more sensitive than conventional RT-PCR and we detected two samples undetected by conventional RT-PCR. The one-step SYBR Green real-time quantitative RT-PCR is specific, sensible and reproducible, which makes it a powerful tool in both diagnostic applications and general research of ARAV and RIOMV and possibly other Brazilian hantaviruses.

show abstract

Detection of hantaviruses in Brazilian rodents by SYBR-Green-based real-time RT-PCR

Cited by 12 publications

References 16 publications

Hantavirus Reservoirs: Current Status with an Emphasis on Data from Brazil

Hantavirus Reservoirs: Current Status with an Emphasis on Data from Brazil

Establishment of SYBR green-based qPCR assay for rapid evaluation and quantification for anti-Hantaan virus compounds in vitro and in suckling mice

Development of a One-Step SYBR Green I Real-Time RT-PCR Assay for the Detection and Quantitation of Araraquara and Rio Mamore Hantavirus

Contact Info

Product

Resources

About