scholarly journals Widespread circulation of echovirus 6 causing aseptic meningitis in paediatric patients in the area of Modena, Italy, in 2011

2016 ◽  
Vol 31 (3) ◽  
Author(s):  
Sara Tagliazucchi ◽  
Francesca Frascaro ◽  
Giulia Fregni Serpini ◽  
Nadia Nanni ◽  
Giulia Forbicini ◽  
...  
Keyword(s):  
Aseptic Meningitis ◽  
Molecular Detection ◽  
Human Enterovirus ◽  
Rt Pcr ◽  
Paediatric Patients ◽  
Emilia Romagna Region ◽  
Polymerase Chain ◽  
Echovirus 6 ◽  
Species Specific ◽  
Information Database

<p><em>Introduction</em>: Between May and November 2011, enterovirus RNA was detected in the cerebrospinal fluids (CSFs) of 72 children with signs of aseptic meningitis admitted to paediatric departments of different Hospitals of the prefecture of Modena, Emilia Romagna region, Italy. Enterovirus RNA was detected in 34 CSFs by commercial reverse transcriptase-polymerase chain reaction (RT-PCR). Twenty-one samples, resulted human enterovirus B by species-specific RT-nested PCR, were submitted to sequencing of the 3’ terminus of the VP1 gene. <br /><em>Materials and Methods:</em> Upon sequencing and interrogation of the National Center for Biotechnology Information database, all 21 viruses were characterized as echovirus 6 (E6), and posses a 100% nucleotide identity each other.<br /><em>Results</em>: This study reports the molecular detection and typing of E6 isolated from clinical specimens from paediatric patients with aseptic meningitis in the wide area of Modena, Italy, in 2011.</p>

scholarly journals Molecular detection and characterisation of mumps virus in cerebrospinal fluid in a Gauteng laboratory

2016 ◽  
Vol 31 (1) ◽  
pp. 29-31
Author(s):  
Marieke Brauer ◽  
Marianne Wolfaardt ◽  
Lynne M. Webber ◽  
Maureen B. Taylor
Keyword(s):  
Polymerase Chain Reaction ◽  
Phylogenetic Analysis ◽  
Cerebrospinal Fluid ◽  
Aseptic Meningitis ◽  
Reverse Transcription ◽  
Molecular Detection ◽  
Mumps Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

The study aimed to determine the presence of mumps virus (MuV) in cerebrospinal fluid (CSF) specimens and to genetically characterise detected MuV strains. A real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the MuV F gene, and characterisation was performed by sequencing of the SH gene. Mumps virus was detected in 1.2% (3/260) of specimens. Phylogenetic analysis of one MuV strain revealed that it clustered with the Jeryl-Lynn and RIT4385 vaccine strains. As far as the authors could ascertain this is the first study to provide viral proof that these vaccine-like strains may be associated with aseptic meningitis.

A Multiplex Reverse Transcription Polymerase Chain Reaction Method for the Detection of Foodborne Viruses†

1999 ◽  
Vol 62 (10) ◽  
pp. 1210-1214 ◽  
Author(s):  
SORAYA I. ROSENFIELD ◽  
LEE-ANN JAYKUS
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Hepatitis A Virus ◽  
Simultaneous Detection ◽  
Polymerase Chain Reaction Method ◽  
Human Enterovirus ◽  
Norwalk Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

A multiplex reverse transcription polymerase chain reaction (RT-PCR) method was developed for the simultaneous detection of the human enteroviruses, hepatitis A virus (HAV) and Norwalk virus (NV). Poliovirus type 1 (PV1) was chosen as a model for the human enterovirus group. Three different sets of primers were used to produce three size-specific amplicons of 435 bp, 270 bp, and 192 bp for PV1, NV, and HAV, respectively. RT-PCR products were separated by agarose gel electrophoresis, and amplicon identity was confirmed by Southern transfer followed by DNA hybridization using nonradio-active, digoxigenin-labeled internal probes. When tested on mixed, purified virus suspensions, the multiplex method achieved detection limits of ≤1 infectious unit (PV1 and HAV) or RT-PCR-amplifiable unit (NV) for all viruses. With further streamlining efforts such as single tube amplification and liquid hybridization, multiplex PCR offers advantages over cell culture methodology and monoplex PCR because it allows for rapid and cost-effective detection of several human enteric viruses in a single reaction tube.

Investigation of Rotavirus infection in sheep usingserological and molecular techniques

2016 ◽  
Author(s):  
Volkan Yilmaz. ◽  
M.Ozkan Timurkan ◽  
Nuvit Coskun ◽  
Yakup Yildirim
Keyword(s):  
Molecular Detection ◽  
Molecular Techniques ◽  
Enzyme Linked Immunosorbent Assay ◽  
Small Scale ◽  
Family Farms ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Rotavirus Infection ◽  
Scale Family ◽  
Polymerase Chain

In this study, serological and molecular research was conducted on the Rotavirus infection in domestic breeds of sheep at 2–3 years of age. The sheep included in the study were raised on small scale family units of less than 20 sheep per unit, in central Kars province and its districts (Susuz, Arpaçay, Kagizman and Selim) in the Northeast Anatolia region of Turkey. The blood and fecal samples were collected randomly from 450 sheep. They were analyzed for the presence of Rotavirus and the antibody against the virus using enzyme-linked immunosorbent assay (ELISA). The highest seropositive ratio (73.46%) was found in central Kars province. The seroprevalence of Rotavirus in sheep raised in the Kars region was determined to be 55.33%. Rotavirus was not detected in fecal samples with ELISA. Molecular detection of Rotavirus from fecal samples was done by reverse transcription polymerase chain reaction (RT-PCR) technique using specific generic primers for VP6 protein. Rotavirus could not be detected in RT-PCR. The data that were obtained showed that the infection spreads on small scale family farms. Based on this information, recommendations were made for controlling Rotavirus infection.

scholarly journals Detection of enteroviruses in cases of neurological disorders in the State of Pará, Brazil

2001 ◽  
Vol 43 (6) ◽  
pp. 321-324 ◽  
Author(s):  
Maria de Lourdes Contente GOMES ◽  
Helena KOPECKA ◽  
Alexandre da Costa LINHARES
Keyword(s):  
Aseptic Meningitis ◽  
Deficiency Syndrome ◽  
Rt Pcr ◽  
Specific Primers ◽  
Newborn Mice ◽  
Negative Results ◽  
Viral Particles ◽  
Motor Deficiency ◽  
Virology Section ◽  
Polymerase Chain

Eighty-one cerebrospinal fluid (CSF) samples mainly from cases of aseptic meningitis and motor deficiency syndrome were sent to the Virology Section of Evandro Chagas Institute, Belém Pará, in the period of January 1995 to January 1996 in order to isolate viruses. All samples were inoculated onto HEp-2 cell culture and newborn mice, with negative results. The probability of isolating viruses by these methods is reduced because of the low concentration of viral particles in these specimens. In order to obtain more information about the etiology of these cases, a group of 23 samples were selected to be tested by a more sensitive technique than the virus isolation - the reverse transcription polymerase chain reaction (RT-PCR). Specific primers directed to conserved regions in the enterovirus genome were used, considering that this group of viruses is frequently associated with these neurological disorder. The age of the patients ranged from 1 to 55 years and nearly all of them lived in Belém, State of Pará, North of Brazil. Of 15 samples analyzed by RT PCR nine (60%) were positive; of these, 6 (66.6%) had motor deficiency and 3 (33.3%) developed aseptic meningitis. These results show that it is important to investigate enterovirus as cause of these syndromes.

Molecular Detection of Tobacco rattle virus in Bleeding Heart [Dicentra spectabilis (L.) Lem.] Growing in Alaska

2013 ◽  
Vol 14 (1) ◽  
pp. 57
Author(s):  
Nancy L. Robertson
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Detection ◽  
Tobacco Rattle Virus ◽  
Virus Transmission ◽  
Rt Pcr ◽  
Chain Reaction ◽  
South Central ◽  
Polymerase Chain ◽  
First Time ◽  
Dicentra Spectabilis

Tobacco rattle virus (TRV) was detected in bleeding heart from South Central Alaskan home gardens in 2010-11. TRV M-type and NM-type isolates were confirmed from these symptomatic bleeding heart plants by reverse transcription (RT)-PCR polymerase chain reaction, protein, serological, and virus transmission assays. RNA1 was sequenced from one of the bleeding heart M-type isolates, and the nucleotide identity ranged from 91% to 94% when compared with six TRV isolates from potato, spinach, and alstroemeria. This is the first detection of TRV from D. spectabilis in Alaska. It is also the first time that M- and NM-type isolates have been distinguished from bleeding heart plants. The significance of these findings is that even though TRV infected plants containing NM-type isolates probably will not be spread to other plants by its specific nematode vector; vegetative propagated roots from TRV infected plants of either type of isolates will continue to be a source of diseased plants to home gardeners. Accepted for publication 18 December 2012. Published 27 February 2013.

scholarly journals Extraction and molecular detection of viral dsRNA from different infected plants

2017 ◽  
Vol 1 ◽  
pp. 197
Author(s):  
Afsaneh Delpasand Khabbazi ◽  
Nemat Sokhandan Bashir ◽  
Saber Delpasand Khabbazi ◽  
Hakimeh Ighani
Keyword(s):  
Molecular Detection ◽  
Extraction Efficiency ◽  
Extraction Procedure ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Viral Dsrna ◽  
Double Stranded Rna ◽  
Modified Method ◽  
Phenol Treatment ◽  
Polymerase Chain

Extraction of viral double stranded RNA (dsRNA) from infected plants is helpful in identification of the viruses involved in infection. To date, there have been several methods developed to isolate dsRNA; however, type of the plant and virus is determinative in extraction efficiency. In this study we extracted dsRNA from different woody and herbaceous plants through a modified method which reduces the costs and time of extraction procedure. This method is based on different affinity of nucleic acids for the cellulose CF-11 in1X STE (Sodium chloride Tris EDTA) buffer containing 16 % ethanol. There is no phenol treatment or mini columns used in the isolation procedure. Extracted dsRNAs were identified by ribonuclease treatment and RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction). We have applied the procedure on five different hosts representing Amaranthaceae, Vitaceae, Fabaceae and Rosaceae infected with four different viruses representing Secoviridae and Bromoviridae. 

scholarly journals Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses

2006 ◽  
Vol 87 (1) ◽  
pp. 119-128 ◽  
Author(s):  
M. Steven Oberste ◽  
Kaija Maher ◽  
Alford J. Williams ◽  
Naomi Dybdahl-Sissoko ◽  
Betty A. Brown ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Pcr Primers ◽  
Human Enterovirus ◽  
Rt Pcr ◽  
Specific Primer ◽  
Specific Primers ◽  
Typical Application ◽  
Partial Sequencing ◽  
Specific Pcr ◽  
Species Specific

The 65 serotypes of human enteroviruses are classified into four species, Human enterovirus (HEV) A to D, based largely on phylogenetic relationships in multiple genome regions. The 3′-non-translated region of enteroviruses is highly conserved within a species but highly divergent between species. From this information, species-specific RT-PCR primers were developed that can be used to rapidly screen collections of enterovirus isolates to identify species of interest. The four primer pairs were 100 % specific when tested against enterovirus prototype strains and panels of isolates of known serotype (a total of 193 isolates). For evaluation in a typical application, the species-specific primers were used to screen 186 previously uncharacterized non-polio enterovirus isolates. The HEV-B primers amplified 68·3 % of isolates, while the HEV-A and HEV-C primers accounted for 9·7 and 11·3 % of isolates, respectively; no isolates were amplified with the HEV-D primers. Twelve isolates (6·5 %) were amplified by more than one primer set and eight isolates (4·3 %) were not amplified by any of the four primer pairs. Serotypes were identified by partial sequencing of the VP1 capsid gene, and in every case sequencing confirmed that the species-specific PCR result was correct; the isolates that were amplified by more than one species-specific primer pair were mixtures of two (11 isolates) or three (one isolate) species of viruses. The eight isolates that were not amplified by the species-specific primers comprised four new serotypes (EV76, EV89, EV90 and EV91) that appear to be unique members of HEV-A based on VP1, 3D and 3′-non-translated region sequences.

scholarly journals DETEKSI MOLEKULER PENYEBAB PENYAKIT KUNING (Tomato chlorosis virus DAN Tomato infectious chlorosis virus) PADA TANAMAN TOMAT (MOLECULAR DETECTION CAUSED YELLOWING DISEASE (Tomato chlorosis virus AND Tomato infectious chlorosis virus) ON TOMATOES)

2017 ◽  
Vol 19 (2) ◽  
pp. 80
Author(s):  
Resti Fajarfika ◽  
Sedyo Hartono ◽  
Sri Sulandari ◽  
Susamto Somowiyarjo
Keyword(s):  
Molecular Detection ◽  
Nucleotide Sequence Analysis ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Pcr Product ◽  
Yellowing Disease ◽  
Polymerase Chain ◽  
Tomato Chlorosis Virus ◽  
Tomato Infectious Chlorosis Virus

ABSTRACTThis research was aimed to detect the ToCV and TICV caused yellowing disease on tomatoes by molecular detection. Leaf samples of symptomatic plants were taken from Ketep (Magelang), then the leaves were identified by reversetranscription-polymerase chain reactions (RT-PCR) using specific primer ToCV-CF/ToCV-CR (360 bp) and TICVCF/TICV-CR (416 bp). The result of nucleotide sequence analysis, amino acid and PCR product phylogenetic sequences were verified as TICV, it showed that TICV from Magelang belongs to the same group with TICV from Japan, North America and Europe, France, Italy, and USA.Keywords: molecular detection, ToCV, TICVINTISARIPenelitian ini bertujuan untuk mendeteksi keberadaan ToCV dan TICV penyebab penyakit kuning pada tanaman tomat. Daun bergejala diambil dari Desa Ketep (Magelang), selanjutnya diuji denganreverse transcription-polymerasechain reactions(RT-PCR) menggunakan primer spesifik ToCV-CF/ToCV-CR (360 bp) dan TICV-CF/TICV-CR (416 bp). Hasil analisis sekuen nukleotida, asam amino, dan filogenetik produk PCR teridentifikasi sebagai TICV yang menunjukkan bahwa TICV isolat Magelang berada dalam satu kelompok dengan isolat TICV asal Jepang, Amerika Utara dan Eropa,Perancis, Italia, dan USA.Kata kunci: deteksi molekuler, ToCV, TICV

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