scholarly journals Molecular Characterization of Echovirus 30 Isolated from Environment in North India

2020 ◽  
Author(s):  
Sarika Tiwari ◽  
Tapan N Dhole
Keyword(s):  
Aseptic Meningitis ◽  
The United States ◽  
North India ◽  
Rt Pcr ◽  
Specific Primers ◽  
Virus Stock ◽  
Echovirus 30 ◽  
Rd Cells ◽  
Infected Cells

Abstract BackgroundEchovirus 30 (E30) causes acute aseptic meningitis. Enteroviruses (EVs) are responsible for 30,000 to 50,000 hospitalizations for aseptic meningitis per year in the United States. E30 is one of the most frequently isolated EVs, causing extensive outbreaks in temperate climates in several countries. MethodsE30 used in this study was isolated from environmental specimens. The virus was confirmed by RT-PCR with specific primers (1). Virus stock was prepared by infecting RD cells in 25 cm2 flasks (Corning Inc. USA). Virus infected and mock infected cells were incubated at 36.5 ºC with 2% MEM. After 48 h of infection, the culture was aliquoted and kept at-80 ºC for further use.ResultsThis present work analyzed the E30 genetic diversity in a fragment of 217 nucleotides of the VP1 region, of environmental strains. The environmental samples of echovirus 30 show mutation in the nonfunctional polyprotein protein (Fig. 1). The result suggested that the information may be obtained by analyzing only a part of the VP1 region.ConclusionThe present study showed that the E30 environmental sample is more divergent to prototype Bastianni strain.

scholarly journals Genetic characterization of a novel recombinant echovirus 30 strain causing a regional epidemic of aseptic meningitis in Hokkaido, Japan, 2017

2019 ◽  
Vol 165 (2) ◽  
pp. 433-438
Author(s):  
Masahiro Miyoshi ◽  
Akiko Goto ◽  
Rika Komagome ◽  
Hiroki Yamaguchi ◽  
Yuji Maruo ◽  
...  
Keyword(s):  
Aseptic Meningitis ◽  
Genetic Characterization ◽  
Echovirus 30

Torovirus detection in faecal specimens of calves and pigs in Hungary: Short communication

2002 ◽  
Vol 50 (3) ◽  
pp. 293-296 ◽  
Author(s):  
Katalin Matiz ◽  
S. Kecskeméti ◽  
I. Kiss ◽  
Keyword(s):  
Electron Microscopy ◽  
Short Communication ◽  
The United States ◽  
Aetiological Agent ◽  
Rt Pcr ◽  
Weaned Piglets ◽  
Chain Reaction ◽  
Specific Primers ◽  
Porcine Torovirus ◽  
Polymerase Chain

Bovine torovirus is an established aetiological agent of disease in cattle, while porcine torovirus has only been isolated from healthy animals. Evidence for the presence of torovirus has been described in several European countries and also in the United States. A survey was performed to detect toroviruses in Hungary by means of sampling ten swine and nine bovine herds. Rectal swabs and faecal specimens were collected from diarrhoeic calves and from weaned piglets. The samples were tested by the reverse transcription-polymerase chain reaction (RT-PCR) using torovirus-specific primers and the positive samples were further examined by electron microscopy (EM). Torovirus was detected in 4 diarrhoeic calves (out of 111) and in 10 healthy weaned pigs (out of 200 tested), representing two of the 9 calf herds and two of the 10 pig herds tested. This is the first report of exact diagnosis of torovirus in Hungary.

scholarly journals Identification and Phylogenetic Characterization of Human Enteroviruses Isolated from Cases of Aseptic Meningitis in Brazil, 2013–2017

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 690 ◽  
Author(s):  
Emanuelle Ramalho ◽  
Ivanildo Sousa ◽  
Fernanda Burlandy ◽  
Eliane Costa ◽  
Amanda Dias ◽  
...  
Keyword(s):  
Aseptic Meningitis ◽  
Neurological Diseases ◽  
Seasonal Pattern ◽  
Temporal Distribution ◽  
Phylogenetic Characterization ◽  
Echovirus 30 ◽  
Sporadic Cases ◽  
Cell Culture Isolation ◽  
Enterovirus Serotypes

Aseptic meningitis is a common viral infection associated with human enteroviruses. The aim of the present study was to identify and characterize the enteroviruses associated with outbreaks and sporadic cases of aseptic meningitis that occurred in different regions of Brazil between 2013 and 2017. Cerebrospinal fluids obtained from patients admitted to public health facilities were analyzed. A total of 303 patients were positive for Human Enteroviruses (EV) by cell culture isolation with a median isolation rate throughout the year of 12%. We were able to identify enterovirus serotypes in 295 clinical specimens. Nineteen different serotypes were identified; the large majority corresponded to HEV-B species. Echovirus 30 (E-30) and Echovirus 6 (E-6) were the most prevalent genotypes (66.8%). Sequence analysis suggested that circulating E-30 was closely related to E-30 from other American countries; while E-6 was derived from Europe. Most of the patients consisted of children ≤ 15 years old. The temporal distribution of all aseptic meningitis and EV-positive cases showed an obvious seasonal pattern during autumn. Our results have provided valuable information about the enteroviral etiology of the aseptic meningitis cases in Brazil pointing to the importance of enterovirus surveillance in neurological diseases.

scholarly journals Karakterisasi Molekuler Papaya ringspot virus tipe P pada Tanaman Mentimun di Jawa

2018 ◽  
Vol 14 (3) ◽  
pp. 75
Author(s):  
Listihani Listihani ◽  
Tri Asmira Damayanti ◽  
Sri Hendrastuti Hidayat ◽  
Suryo Wiyono
Keyword(s):  
Ringspot Virus ◽  
Mosaic Symptom ◽  
Papaya Ringspot Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Specific Primers ◽  
Central Java ◽  
Dna Fragment ◽  
Polymerase Chain

Moleculer Characterization of Papaya ringspot virus type P on Cucumber in JavaInfection of Papaya ringspot virus (PRSV) on cucumber plants showing mosaic symptom was detected using specific antibody.  Further investigation was conducted to determine molecular characters and status of PRSV infecting cucumber in Java.  Infection of PRSV was detected from leaf samples collected from the field using dot immunobinding assay (DIBA).  Disease frequency caused by PRSV infection reached 81.11%, 95.86%, 91.66%, and 92.3% in East Java, Central Java, Yogyakarta, and West Java, respectively.  Characterization of PRSV isolates was conducted by reverse transcription polymerase chain reaction (RT-PCR) using specific primers for PRSV-P and PRSV-W, followed by cloning, and DNA sequencing.  DNA fragment of 470 bp was successfully amplified using specific primers for PRSV-P from several samples from Nganjuk, Brebes, Kulon Progo, and Subang; but no amplification was achieved using specific primers for PRSV-W.  Nucleotide and amino acid analysis showed high homology among PRSV-P isolates from Nganjuk, Brebes, Kulon Progo, and Subang, i.e. 98.6%-99.7% and 99.3%-100%, respectively.  This is an indication of a low genetic variation among PRSV-P from Java. Further phylogenetic analysis indicated that PRSV-P isolate cucumber is in the same cluster with PRSV-P isolate papaya from Bali, Indonesia.  This is the first report of PRSV-P infecting cucumber in Indonesia.

scholarly journals First Report of Tomato chlorosis virus and Tomato infectious chlorosis virus in Tomato Crops in France

Plant Disease ◽  
2005 ◽  
Vol 89 (11) ◽  
pp. 1243-1243 ◽  
Author(s):  
A. Dalmon ◽  
S. Bouyer ◽  
M. Cailly ◽  
M. Girard ◽  
H. Lecoq ◽  
...  
Keyword(s):  
Protein Gene ◽  
The United States ◽  
Rt Pcr ◽  
Specific Primers ◽  
Southern France ◽  
Sequence Identity ◽  
Two Samples ◽  
Dna Fragment ◽  
Tomato Chlorosis Virus ◽  
Tomato Infectious Chlorosis Virus

Since 2002, yellowing symptoms associated with high levels of white-fly populations have been observed in plants of protected tomato crops in France. Symptomatic plants exhibited interveinal yellowing areas in older leaves, followed by generalized yellowing. Symptoms were not observed in young plants or fruits. Trialeurodes vaporariorum populations were generally abundant in spring, and Bemisia tabaci (established in France for approximately 10 years) became predominant in summer and fall. To check for the presence of Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV), two whitefly-transmitted criniviruses known to induce yellowing symptoms, 696 samples were collected in the major tomato-growing areas; 573 samples from southern France and 123 samples from northern France. Total RNA was extracted from each sample and analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Primers specific to ToCV (2) and TICV (1,3) were used to amplify either part of the heat-shock-like protein gene HSP70h (both viruses) or part of the diverged coat protein gene (CPd), (TICV only). A 439-bp DNA fragment was obtained with ToCV primers in 178 samples from southern France collected mainly from mid-spring to early fall from 2002 to 2004. Three RT-PCR products amplified from samples collected from diverse growing areas were sequenced and showed 99 to 100% sequence identity with published ToCV sequences from Spain (GenBank Accession Nos. AF215818, AF233435, and AF215817), Portugal (GenBank Accession No. AF234029), Sicily (GenBank Accession No. AY048854), and the United States (GenBank Accession No. AF024630). Considering the high frequency of ToCV-infected samples (41 positive samples of 112 samples collected in 2002, 71 of 295 collected in 2003, and 66 of 166 collected in 2004), this virus appears to be well established in southern France but remains absent in the northern regions. The presence of TICV was tested in 485 samples using the CPd-specific primers or the HSP70h-specific primers. The virus was detected in only two samples from Nice (southeastern France) in 2003 with both primer pairs. The CPd DNA fragment (700 bp) from one of these samples was sequenced, showing 98.9% sequence identity with a TICV Japanese isolate (AB085603). Results of these assays suggest that in contrast to ToCV, TICV is not yet broadly established in France. This difference could be associated with the specificity of the vectors, since ToCV is transmitted by B. tabaci and T. vaporariorum, while TICV is transmitted only by T. vaporariorum (4). References: (1) R. H. Li et al. Plant Dis. 82:84, 1998. (2) D. Louro et al. Eur. J. Plant Pathol. 1065:589, 2000. (3) A. M. Vaira et al. Phytoparasitica 30:290, 2002. (4) G. C. Wisler et al. Plant Dis. 82:271, 1998.

scholarly journals First Report of Rose rosette virus Associated with Rose Rosette Disease Infecting Knockout Roses in Florida

Plant Disease ◽  
2014 ◽  
Vol 98 (10) ◽  
pp. 1449-1449 ◽  
Author(s):  
B. Babu ◽  
H. Dankers ◽  
E. Newberry ◽  
C. Baker ◽  
T. Schubert ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Pairwise Comparison ◽  
The United States ◽  
Rt Pcr ◽  
Specific Primers ◽  
Knock Out ◽  
First Report ◽  
Plant Pathol ◽  
Buffer Control ◽  
Rose Rosette

Roses are one of the most popular flowering shrubs in the United States, with a total wholesale value of US$194 million. Among the major states, Florida is the fourth largest producer of roses with a total value exceeding US$20 million (4). In Florida, the roses have become especially popular in recent years with the introduction of Knock Out and other shrub roses. Virus-like symptoms including witches'-broom, excessive thorns, abnormal red discoloration of shoots and foliages, distorted leaves, and deformed buds and flowers were initially observed on Knock Out roses in a commercial nursery in Quincy, FL, in November 2013. Fifteen plants out of ~250,000 plants showed these characteristic symptoms. Total RNA extracts (RNeasy Plant Mini Kit, Qiagen, Valencia, CA) from eight symptomatic and two non-symptomatic rose samples were subjected to reverse-transcription (RT) assays using SuperScript III Reverse transcriptase (Invitrogen, Life Technologies, NY) and random hexamer primers. The cDNA synthesized was then subjected to PCR assay using Platinum Taq DNA polymerase (Invitrogen, Life Technologies) and using Rose rosette virus (RRV) specific primers RRV-F and RRV-R (1), targeting the core region of the RNA1 genome of the virus. The RT-PCR assays using the specific primers produced amplicons of 375 bp, only in the symptomatic leaf samples. The obtained amplicons were PCR purified and sequenced directly (GenBank Accession Nos. KF990370 to KF990377). BLAST analysis of these sequences revealed a higher identity of 99% with the RRV (HQ871942) in the NCBI database. Pairwise comparison of the eight RRV sequences exhibited 99 to 100% identity among themselves. These results revealed the association of RRV with the symptomatic rose plants. Eight symptomatic and two non-symptomatic rose plant samples were tested for RRV using blot hybridization assay, utilizing a digoxigenin-labeled DNA probe of 511 bp, targeting the RNA1 genome of the RRV. All eight symptomatic rose plants showed a positive reaction to the RRV-specific probes, confirming the presence of RRV in the samples, while the non-symptomatic and the buffer control did not produce any reactions. Even though the virus is reported to spread by an eriophyid mite Phyllocoptes fructiphilus, thorough examination of the infected samples showed absence of the vector. The samples were also tested using RT-PCR for the presence of Rose cryptic virus (RCV) and Blackberry chlorotic ringspot virus (BCRV) using specific primers (2,3). The samples tested negative for the RCV and BCRV. This is the first report of occurrence of RRV on rose in Florida. Considering the economic importance of the rose plants and the highly destructive nature of RRV, this report underscores the need for immediate effective quarantine and management of the virus for protecting the economically important rose industry in Florida. References: (1) A. G. Laney et al. J. Gen. Virol. 92:1727, 2011. (2) S. Sabanadzovic and N. Abou Ghanem-Sabanadzovic. J. Plant Pathol. 90:287, 2008. (3) I. E. Tzanetakis et al. Plant Pathol. 55:568, 2006. (4) USDA. 2007 Census of Agriculture 3:25, Washington, DC, 2010.

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.

scholarly journals Detection by PCR of Enteroviruses in Cerebrospinal Fluid during a Summer Outbreak of Aseptic Meningitis in Switzerland

1998 ◽  
Vol 36 (9) ◽  
pp. 2408-2412 ◽  
Author(s):  
Meri Gorgievski-Hrisoho ◽  
Jean-Daniel Schumacher ◽  
Nevenka Vilimonovic ◽  
Daniel Germann ◽  
Lukas Matter
Keyword(s):  
Cell Culture ◽  
Cerebrospinal Fluid ◽  
Aseptic Meningitis ◽  
Diagnostic Techniques ◽  
Rt Pcr ◽  
Echovirus 30 ◽  
Reverse Transcription Pcr ◽  
Viral Culture ◽  
Test Kit ◽  
Virus Culture

Enteroviruses (EV) are among the most common causes of aseptic meningitis. Standard diagnostic techniques are often too slow and lack sensitivity to be of clinical relevance. EV RNA can be detected within 5 h by a commercially available reverse transcription-PCR (RT-PCR) test kit. Cerebrospinal fluid (CSF) samples from 68 patients presenting with aseptic meningitis during a summer outbreak in Switzerland were examined in parallel with cell culture and commercial RT-PCR. RT-PCR was positive in all 16 CSF specimens positive by cell culture (100%). In addition, 42 of 52 (80%) CSF samples negative by cell culture were PCR positive. In 26 of these 42 (62%) patients, viral culture from other sites (throat swab or stool) was also positive. The CSF virus culture took 3 to 7 days to become positive. Echovirus 30 was the type most often isolated in this outbreak. The sensitivity of CSF RT-PCR based on clinical diagnosis during this aseptic meningitis outbreak in patients with negative bacterial culture results was 85%, i.e., considerably higher than the sensitivity of CSF virus culture (24%). We conclude that this commercial RT-PCR assay allows a positive diagnosis with minimal delay and may thus influence clinical decisions.

scholarly journals Retrospective characterization of Newcastle Disease Virus Antrim '73 in relation to other epidemics, past and present

2004 ◽  
Vol 132 (2) ◽  
pp. 357-368 ◽  
Author(s):  
K. O'DONOGHUE ◽  
B. LOMNICZI ◽  
B. McFERRAN ◽  
T. J. CONNOR ◽  
B. SEAL ◽  
...  
Keyword(s):  
Northern Ireland ◽  
Latin American ◽  
Newcastle Disease ◽  
Western Europe ◽  
Phylogenetic Analyses ◽  
The United States ◽  
The Other ◽  
Rt Pcr ◽  
Disease Free

In November 1973 Newcastle disease suddenly appeared in Northern Ireland, where the viscerotropic disease had not been seen in 3½ years and the two Irelands had been regarded as largely disease free for 30 years. It was successfully controlled with only 36 confirmed affected layer flocks, plus 10 more slaughtered as ‘dangerous contacts’. Contemporary investigations failed to reveal the source of the Irish epidemic. Using archival virus samples from most of the affected flocks, RT–PCR was conducted with primers selected for all six NDV genes. Phylogenetic analyses of three genes, HN, M and F, confirmed vaccine as the cause of one of the outbreaks. The other six samples were identical and closely related to previous outbreaks in the United States and western Europe initiated by infected imported Latin American parrots. The probable cause of the epidemic followed from the importation from The Netherlands of bulk feed grains contaminated with infected pigeon faeces.

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