scholarly journals Absence of paramyxovirus RNA in non-human primate sanctuaries and a primatology center in Gabon

2019 ◽  
Vol 5 (2) ◽  
pp. 6 ◽  
Author(s):  
Barthelemy Ngoubangoye ◽  
Gael Darren Maganga ◽  
Larson Boundenga ◽  
Thierry Audrey Tsoumbou ◽  
Virginie Rougeron ◽  
...  
Keyword(s):  
Medical Research ◽  
Reverse Transcription ◽  
Host Species ◽  
Conservation Programs ◽  
Potential Host ◽  
Reverse Transcription Pcr ◽  
Circulation Patterns ◽  
Wide Range ◽  
Species Groups ◽  
Human Primate

The viruses of the Paramyxoviridae family are known to infect a wide range of animals, including primates, birds, rodents,carnivores, bats, ungulates, snakes, cetaceans and humans. This study aims to investigate the circulation of paramyxoviruses in five potential host species groups (humans, non-human primates, rodents, shrews, and bats) living in the same environments in three conservation programs dedicated to non-human primates, namely the Lékédi park, the primatology center of the International Center for Medical Research of Franceville and the Gorilla Protection Program, located in Gabon. We tested 35 workers, 343 NHPs (8 species), 141 bats (4 species), 420 rodents (5 species) and 10 shrews, sampled between 2013 and 2014. Faecal and organ samples were analyzed using three heminested reverse transcription-PCR (hnRT-PCR). All the 1884 samples tested were negative for PV detection. Further studies spanning a greater period of time are needed to investigate PV circulation patterns in theseconservation programs.

scholarly journals COVID-19 in children in Odisha state, India: a retrospective review

2021 ◽  
Vol 5 (1) ◽  
pp. e001284
Author(s):  
Girish Chandra Dash ◽  
Subhra Subhadra ◽  
Jyotirmayee Turuk ◽  
Debaprasad Parai ◽  
Usha Kiran Rout ◽  
...  
Keyword(s):  
Young People ◽  
Medical Research ◽  
Reverse Transcription ◽  
Retrospective Review ◽  
Research Centre ◽  
Children And Young People ◽  
Indian Council ◽  
Reverse Transcription Pcr ◽  
Regional Medical ◽  
Regional Medical Research

We retrospectively analysed the swab samples tested for COVID-19 from 7 March 2020 to 17 August 2021 at the Indian Council of Medical Research-Regional Medical Research Centre, Bhubaneswar, Odisha. 553 763 nasopharyngeal swabs were collected from individuals suspected with COVID-19 in Odisha state. 75 190 (13.6%) samples were positive by reverse transcription-PCR. There were 5988 (8%) cases in children and young people under 18 years old. Odisha reported 996 153 COVID-19 cases which resulted in 6985 deaths in adults and 36 in children and young people under 18 years old.

scholarly journals Cytochrome P450 1B1 mRNA measured in blood mononuclear cells by quantitative reverse transcription-PCR

1998 ◽  
Vol 44 (12) ◽  
pp. 2416-2421 ◽  
Author(s):  
Cristina Dassi ◽  
Stefano Signorini ◽  
Piermario Gerthoux ◽  
Mariangela Cazzaniga ◽  
Paolo Brambilla
Keyword(s):  
Cytochrome P450 ◽  
Aromatic Hydrocarbons ◽  
Reverse Transcription ◽  
Mononuclear Cells ◽  
Internal Standard ◽  
Reverse Transcription Pcr ◽  
Wide Range ◽  
Two Samples ◽  
Blood Mononuclear Cells ◽  
Cyp1b1 Mrna

Abstract Cytochrome P450 (CYP) 1B1 activates polycyclic aromatic hydrocarbons and aryl aromatic hydrocarbons to carcinogens. We describe a competitive reverse transcription-PCR (RT-PCR) assay for the quantification of CYP1B1 mRNA in blood mononuclear cells (BMCs) by simultaneous RT and PCR amplification of cellular RNA with decreasing amounts of an internal standard. The concentration of CYP1B1 mRNA is derived from the ratio between the intensities of the bands corresponding to the amplified products. To reduce the variability of mRNA extraction efficiency, the measured amount of CYP1B1 has been calculated in relation to the β-actin gene products. We measured CYP1B1 expression in the BMCs of 75 human subjects; no significant differences in the CYP1B1:β-actin ratio were detected between women (range, 0.47–4.35; median, 2.0) and men (range, 0.72–3.85; median, 2.09). The analytical imprecision (CV) of duplicates was 14% (n = 25 pairs), and the intraindividual CV for two samples, 1 month apart, was 22% (n = 20). No significant differences were detected in smokers (n = 25; range, 0.77–3.55; median, 2.14) compared with nonsmokers (n = 50; range, 0.47–4.35; median, 2.0). The method has a wide range of linearity, good sensitivity and precision, and is suitable for studies of individual susceptibility as indicated by CYP1B1 expression in BMCs.

scholarly journals Identification of Salt-Stress-Induced Genes from the RNA-Seq Data ofReaumuria trigynaUsing Differential-Display Reverse Transcription PCR

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Zhen-hua Dang ◽  
Qi Qi ◽  
Hui-rong Zhang ◽  
Hao-yu Li ◽  
Shu-Biao Wu ◽  
...  
Keyword(s):  
Salt Stress ◽  
Differential Display ◽  
Reverse Transcription ◽  
Expression Patterns ◽  
Digital Gene Expression ◽  
Rna Seq ◽  
Sequencing Data ◽  
Reverse Transcription Pcr ◽  
Wide Range ◽  
Differential Display Reverse Transcription

Next generation sequencing (NGS) technologies have been used to generate huge amounts of sequencing data from many organisms. However, the correct choice of candidate genes and prevention of false-positive results computed from digital gene expression (DGE) of RNA-seq data are vital when using these genetic resources. We indirectly identified 18 salt-stress-inducedReaumuria trigynatranscripts from the transcriptome sequencing data using differential-display reverse transcription PCR (DDRT-PCR) combined with local BLAST searches. Highly consistent with the DGE results, the quantitative real-time PCR expression patterns of these transcripts showed strong upregulation by salt stress, suggesting that these genes may play important roles inR. trigyna’s survival under high-salt environments. The method presented here successfully identified responsive genes from the massive amount of RNA-seq data. Thus, we suggest that DDRT-PCR could be employed to mine NGS data in a wide range of applications in transcriptomic studies. In addition, the genes identified in the present study are promising candidates for further elucidation of the salt tolerance mechanisms inR. trigyna.

scholarly journals Genogroup I and II Noroviruses Detected in Stool Samples by Real-Time Reverse Transcription-PCR Using Highly Degenerate Universal Primers

2004 ◽  
Vol 70 (12) ◽  
pp. 7179-7184 ◽  
Author(s):  
Gary P. Richards ◽  
Michael A. Watson ◽  
Rebecca L. Fankhauser ◽  
Stephan S. Monroe
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Rna Extraction ◽  
Universal Primers ◽  
Rt Pcr ◽  
Degenerate Primers ◽  
Reverse Transcription Pcr ◽  
Wide Range ◽  
Genetic Clusters ◽  
Stool Samples

ABSTRACT Genogroup I noroviruses from five genetic clusters and genogroup II noroviruses from eight genetic clusters were detected in stool extracts using degenerate primers and single-tube, real-time reverse transcription-PCR (RT-PCR) with SYBR Green detection. Two degenerate primer sets, designated MON 431-433 and MON 432-434, were designed from consensus sequences from the major clusters of norovirus based on the RNA-dependent RNA polymerase region of the norovirus genome. Viruses were extracted from stool samples within 20 min using a viral RNA extraction kit. Real-time RT-PCR for noroviruses generated semiquantitative results by means of the cycle threshold data and dilution endpoint standard curves. Presumptive product verification was achieved by evaluation of first-derivative melt graphs. Multiple clusters of noroviruses were identified simultaneously in a multiplex fashion by virtue of slight differences in melting temperature. The detection of 13 different genetic clusters suggests that the MON primers may serve as universal primers for most, if not all, of the noroviruses in a multiplex assay. Our technique provides a framework for broad application of real-time RT-PCR in clinical, environmental, and food testing laboratories for a wide range of noroviruses.

scholarly journals Morphometric and Molecular Analyses of Ostertagia leptospicularis Assadov, 1953 from Ruminants: Species Diversity or Host Influence?

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 182
Author(s):  
Anna Wyrobisz-Papiewska ◽  
Jerzy Kowal ◽  
Elżbieta Łopieńska-Biernat ◽  
Paweł Nosal ◽  
Iwona Polak ◽  
...  
Keyword(s):  
Host Species ◽  
Roe Deer ◽  
Bos Taurus ◽  
Capreolus Capreolus ◽  
Nematode Species ◽  
Molecular Techniques ◽  
Systematic Classification ◽  
Morphological Variations ◽  
Wide Range ◽  
Induced Changes

Ostertagia leptospicularis Assadov, 1953 was formally described in roe deer Capreolus capreolus and has been reported in a wide range of ruminants, including other Cervidae, as well as Bovidae. Nematode specimens derived from various host species exhibit morphological similarity; however, some differences can be observed. It is unclear if this is due to the differential reaction of one nematode species in different host species (i.e., host-induced changes) or because of distinct nematode species in these hosts (i.e., species complex). This paper focuses on specimens resembling O. leptospicularis f. leptospicularis and its closely related species (Ostertagia ostertagi f. ostertagi) collected from various hosts. Morphometric and molecular techniques were applied to assess host-induced changes in nematode morphology and to clarify its systematic classification. There was an overall effect of host species on measurements of nematodes resembling O. leptospicularis (both males and females), but the distinctiveness of the specimens from cattle Bos taurus were highlighted. The results obtained may suggest that the specimens of O. leptospicularis from cattle in Germany and cervids in central Europe belong to different strains. Furthermore, nematodes from the cervid strain appear to circulate within particular host species, which can be seen in the stated morphological variations.

scholarly journals Implementation of an in-house real-time reverse transcription-PCR assay to detect the emerging SARS-CoV-2 N501Y variants

2021 ◽  
pp. 104868
Author(s):  
Marielle BEDOTTO ◽  
Pierre-Edouard FOURNIER ◽  
Linda HOUHAMDI ◽  
Philippe COLSON ◽  
Didier RAOULT
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Pcr Assay ◽  
Reverse Transcription Pcr

scholarly journals Reverse Transcription-PCR Analysis of Bottled and Natural Mineral Waters for the Presence of Noroviruses

2003 ◽  
Vol 69 (11) ◽  
pp. 6541-6549 ◽  
Author(s):  
Gilbert Thierry Lamothe ◽  
Thierry Putallaz ◽  
Han Joosten ◽  
Joey D. Marugg
Keyword(s):  
Reverse Transcription ◽  
Membrane Filtration ◽  
Limit Of Detection ◽  
Mineral Waters ◽  
Pcr Analysis ◽  
Rna Sequences ◽  
Natural Mineral ◽  
Reverse Transcription Pcr ◽  
Viral Particles ◽  
Nonspecific Amplification

ABSTRACT A seminested reverse transcription-PCR method coupled to membrane filtration was optimized to investigate the presence of norovirus (NV) RNA sequences in bottled and natural mineral waters. The recovery of viral particles by filtration varied between 28 and 45%, while the limit of detection of the overall method ranged from 6 to 95 viral particles. The assay was broadly reactive, as shown by the successful detection of 27 different viral strains representing 12 common genotypes of NVs. A total of 718 bottled and natural mineral water samples were investigated, including 640 samples of finished, spring, and line products (mostly 1 to 1.5 liters), collected from 36 different water brands of various types and from diverse geographic origins over a 2-year period. In addition, 78 samples of larger volume (10 and 400 to 500 liters) and environmental swabs were investigated. From the 1,436 analyses that were performed for the detection of NVs belonging to genogroups I and II, 34 samples (2.44%) were presumptively positive by seminested RT-PCR. However, confirmation by DNA sequence analysis revealed that all presumptive positive results were either due to nonspecific amplification or to cross-contamination. In conclusion, these results do not provide any evidence for the presence of NV genome sequences in bottled waters.

Revision of Coronostrongylus (Nematoda : Strongyloidea) parasitic in the stomachs of macropodid marsupials

2002 ◽  
Vol 16 (6) ◽  
pp. 893 ◽  
Author(s):  
I. Beveridge
Keyword(s):  
Papua New Guinea ◽  
Host Species ◽  
Related Species ◽  
New Guinea ◽  
Nematode Species ◽  
Closely Related Species ◽  
Nematode Genus ◽  
Eastern Australia ◽  
Wide Range ◽  
Species Occurrences

The monotypic nematode genus Coronostrongylus Johnston & Mawson, 1939 from the stomachs of macropodid marsupials was reviewed and was found to consist of a least seven closely related species. Coronostrongylus coronatus Johnston & Mawson, 1939 is found most commonly in Macropus rufogriseus, but occurs occasionally in M. dorsalis, M. parryi and Petrogale inornata. Coronostrongylus johnsoni, sp. nov. is most commonly found in M. dorsalis, but occurs also in M. rufogriseus, M. parma, Thylogale stigmatica, Petrogale godmani and P. brachyotis. Coronostrongylus barkeri, sp. nov. is most prevalent in Onychogalea unguifera, but occurs also in M. rufus, M. robustus and P. brachyotis. Coronostrongylus closei, sp. nov. is restricted to Petrogale persephone. Coronostrongylus sharmani, sp. nov. occurs only in rock wallabies from eastern Australia: P.�coenensis, P. godmani and P. mareeba; C. spratti, sp. nov. occurs in P. inornata and P. assimilis. Coronostrongylus spearei, sp. nov. is restricted to Papua New Guinea where it is found in Dorcopsulus vanhearni, Dorcopsis hageni and D. muelleri. Although all of the nematode species occur in one principal host species or a series of closely related host species, occurrences in geographically disjunct areas and in phylogenetically distant hosts are features of C. coronatus, C. barkeri, sp. nov. and C. johnsoni, sp. nov. The occurrence of seven closely related nematode species found in a wide range of macropodid host species is more readily accounted for by a hypothesis involving multiple colonisations of hosts than by the hypothesis of co-speciation.

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