scholarly journals Serological and Molecular Characterization of Avian Metapneumovirus in Chickens in Northern Vietnam

2021 ◽  
Vol 8 (10) ◽  
pp. 206
Author(s):  
Van-Giap Nguyen ◽  
Hee-Chun Chung ◽  
Hai-Quynh Do ◽  
Thanh-Trung Nguyen ◽  
Thi-Bich-Phuong Cao ◽  
...  
Keyword(s):  
Respiratory Diseases ◽  
Genetic Characterization ◽  
Age Groups ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Avian Metapneumovirus ◽  
Northern Vietnam ◽  
Polymerase Chain

Avian Metapneumovirus (aMPV) is a causative agent of respiratory disease complex in turkeys and chickens that has recently been detected in Vietnam. Due to its novelty, this study was conducted to elucidate the distribution of aMPV in several provinces in northern Vietnam. By the application of Enzyme-Linked Immunosorbent Assay (ELISA) and nested Reverse Transcription-Polymerase Chain Reaction (RT-PCR), this study demonstrated the circulation of aMPV in 12 out of 14 cities/provinces with positive rates of 37.6% and 17.2%, respectively. All nested RT-PCR positive samples were aMPV subgroup B. By pairing the detection results with age groups, it was observed that aMPV infections occurred in chickens of all ages. Additionally, by genetic characterization, aMPV strains were demonstrated to not be attenuated vaccine viruses and to belong to at least two genetic clades. Overall, the obtained results provided insights into the prevalence of aMPV and indicated a greater complexity of respiratory diseases in chickens in Vietnam.

scholarly journals Diagnosis and characterization of canine parvovirus-2 affecting canines of South Gujarat, India

2018 ◽  
Vol 87 (3) ◽  
pp. 247-254
Author(s):  
Kishan Kumar Sharma ◽  
Irsadullakhan Habibullakhan Kalyani ◽  
Shailee Manishbhai Pandya ◽  
Jignesh Alabhai Vala
Keyword(s):  
Age Groups ◽  
Rapid Test ◽  
Canine Parvovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sequencing Analysis ◽  
Nucleotide Polymorphism ◽  
Chain Reaction ◽  
Single Nucleotide ◽  
Polymerase Chain

The present study was carried out in the region of South Gujarat, India, to determine the prevalence and predisposing factors of canine parvovirus-2 (CPV-2) infection in acute gastroenteritis of pups. Further, haemagglutination (HA) test, enzyme linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) and rapid immunochromatography test were compared for diagnosis and prevalent CPV-2 types were ascertained. A total of 73 diarrhoea samples were collected and out of those 32, 33 and 35 were found positive via HA, ELISA and PCR, respectively. In rapid test, 26/52 samples were found positive. Among different age-groups, 11/24 and 13/21 animals were positive in pups aged 4–8 and 8–12 weeks, respectively. All but one (34/35) positive samples were from unvaccinated animals. Labrador was found to be the most susceptible breed (n = 13) to infection. Considering PCR as the best test, 47.94% (35/73) prevalence of CPV was recorded. Among PCR positive samples, 3 and 32 belonged to type CPV-2a and CPV-2b, respectively. Type CPV-2c was not detected among the examined samples. Sequencing analysis of 9/10 CPV-2b isolates revealed single nucleotide polymorphism (SNP) (A-G) at position 4106 (alanine to threonine) and suggested the occurrence of mutant, new CPV-2b in this area. As other major pathogen canine coronavirus was detected in 7/38 CPV negative samples. Conclusively, CPV-2 infection was detected in 47.97% cases of AGE of pups which warrants search for other pathogens in the diagnostic procedure. This work is among the few recent reports which depict the occurrence of a novel mutant (new CPV-2b) in India.

scholarly journals Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Genome ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Kappa Value ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Level Of Agreement

<b><i>Background:</i></b> Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. <b><i>Methods:</i></b> A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. <b><i>Results:</i></b> S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (<i>p</i> = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. <b><i>Conclusions:</i></b> The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

scholarly journals Genetic diversity of Blastocystis in kindergarten children in southern Xinjiang, China

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Meng Qi ◽  
Zilin Wei ◽  
Ying Zhang ◽  
Qiyuan Zhang ◽  
Juanfeng Li ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Genetic Characterization ◽  
Intestinal Parasites ◽  
Small Subunit ◽  
Kindergarten Children ◽  
Chain Reaction ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Southern Xinjiang

Abstract Background Blastocystis is one of the most common intestinal parasites in humans and various animals worldwide. Few studies are available regarding the genetic characterization of Blastocystis infections in humans in China. Methods In the present study, 609 fecal samples were collected from two- to six-year-old kindergarten children in southern Xinjiang and were examined by polymerase chain reaction (PCR). Results The infection rate of Blastocystis was 14.3% (87/609); no significant difference was observed among counties and between sexes. Blastocystis subtypes ST1 (n = 38), ST2 (n = 8), and ST3 (n = 41) were identified by sequence analysis of the small subunit ribosomal RNA gene. Genetic polymorphisms were observed at the intra-subtype level, including seven variations for ST1 (ST1A to ST1G), four for ST2 (ST2A to ST2D), and two for ST3 (ST3A and ST3B); with ST1F and ST2B being new variations. Conclusions ST1 and ST3 are the two common Blastocystis subtypes in the study area. More extensive studies in both humans and animals in different regions are needed to better characterize the transmission of Blastocystis.

Detection of small round structured viruses in clinical and environmental samples by polymerase chain reaction

1995 ◽  
Vol 31 (5-6) ◽  
pp. 375-382 ◽  
Author(s):  
M. Wolfaardt ◽  
C. L. Moe ◽  
W. O. K. Grabow
Keyword(s):  
Polymerase Chain Reaction ◽  
Tap Water ◽  
Practical Method ◽  
Glass Wool ◽  
Polluted Water ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Elution Method

Norwalk (NV) and other small round structured viruses (SRSVs) have been identified as common causes of gastroenteritis. Outbreaks of Norwalk gastroenteritis have been associated with contaminated drinking water and food such as oysters and salads. The cloning and sequencing of the NV genome has made it possible to detect NV and related viruses by the reverse transcription-polymerase chain reaction (RT-PCR). We applied RT-PCR to detect SRSVs in faecal specimens from two gastroenteritis outbreaks in South Africa, designated “Christmas” and “Grootbrak” and were able to detect SRSVs in all of the three specimens from the Christmas outbreak and in two of 16 specimens from the Grootbrak outbreak. The RT-PCR procedure used appeared to be more sensitive for the detection of SRSVs in patient stool specimens than immune electron microscopy and NV antigen detection by enzyme linked immunosorbent assay. The RT-PCR procedure proved suitable for the detection of SRSVs in seeded samples of sewage, sewage sludge, river water, and tap water. However, sensitivity was lower for seeded samples of sewage and sludge than for tap water, which indicates interference by high levels of organic matter. The RT-PCR procedure was also used to show that small numbers of SRSVs can successfully be recovered from large volumes of water by means of a glass wool adsorption-elution method. Since no practical method is available for quantitation of the small numbers of SRSVs concerned, it was not possible to evaluate the efficiency of recovery. Although no SRSVs have been detected by direct testing of sewage and sludge samples, the results obtained in this study show that RT-PCR detection of SRSVs in sewage and polluted water environments is feasible, and that small numbers of the viruses can, like many other enteric viruses, successfully be recovered by means of a glass wool adsorption-elution method.

scholarly journals RT-PCR Method for Detecting Cowpea Mottle Carmovirus in Vigna Germ Plasm

Plant Disease ◽  
1999 ◽  
Vol 83 (7) ◽  
pp. 639-643 ◽  
Author(s):  
A. G. Gillaspie ◽  
S. E. Mitchell ◽  
G. W. Stuart ◽  
R. F. Bozarth
Keyword(s):  
Germ Plasm ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mild Mosaic ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Viral Pathogens ◽  
Highly Sensitive ◽  
Pcr Method ◽  
The Common ◽  
Polymerase Chain

A highly sensitive reverse transcription-polymerase chain reaction (RT-PCR) method was developed to detect cowpea mottle carmovirus (CPMoV) in newly acquired germ plasm of Vigna spp. It detected virus in tissues diluted up to 10-9. The preferred primers were designed from the RNA replicase cDNA sequence of CPMoV. These primers were able to detect CPMoV in plants infected with 10 different isolates of the virus. There were no cross-reactions with either bean mild mosaic or melon necrotic spot carmoviruses or any of the common cowpea viral pathogens tested. The RT-PCR method was up to 105 times more sensitive than direct antigen coating enzyme-linked immunosorbent assay (DAC-ELISA) in detecting CPMoV. The RT-PCR method gave no false positive reaction as is sometimes seen with ELISA.

The Polymerase Chain Reaction (PCR) in the routine genetic characterization of retinoblastoma: A tool for the clinical laboratory

1997 ◽  
Vol 41 (4) ◽  
pp. 331-340 ◽  
Author(s):  
Domenico Mastrangelo ◽  
Nicola Squitieri ◽  
Stefano Bruni ◽  
Theodora Hadjistilianou ◽  
Renato Frezzotti
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Characterization ◽  
Clinical Laboratory ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Circulation of bluetongue virus in goats in the Karamoja region of Uganda

2013 ◽  
Vol 84 (1) ◽  
Author(s):  
Elijah N. Mulabbi ◽  
Chrisostom Ayebazibwe ◽  
Samuel Majalija ◽  
Carrie A. Batten ◽  
Christopher A.L. Oura
Keyword(s):  
Real Time ◽  
Whole Blood ◽  
Bluetongue Virus ◽  
Rna Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
High Seroprevalence ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Karamoja Region

The presence of bluetongue virus (BTV) in indigenous goats from the Karamoja region of northern Uganda was investigated. A total of 300 goats were sampled (serum and whole blood) from five districts within the Karamoja region. The samples were analysed for the presence of bluetongue (BT) antibodies using a commercial Enzyme-linked immunosorbent assay (ELISA) and for the presence of BTV viral RNA by real-time Reverse transcription polymerase chain reaction (RT-PCR), because BTV is an RNA virus. Of the 300 goats tested, 269 (90%) were positive for BTV antibodies, indicating high levels of BTV circulation within the region. Out of the 150 whole blood samples tested for the presence of the virus by real-time RT-PCR, 84 (56%) were positive for BTV RNA. This study, which is the first of its kind in Uganda, showed a high seroprevalence of BT antibodies and active circulation of BTV in a high proportion of goats in the Karamoja region.

scholarly journals First Detection and Characterization of Chrysanthemum virus B infecting Chrysanthemum in Thailand

2021 ◽  
Author(s):  
Salit Supakitthanakorn ◽  
Garnjana Wichitrakoonthavorn ◽  
Kaewalin Kunasakdakul ◽  
On-Uma Ruangwong
Keyword(s):  
Cultural Values ◽  
Ornamental Plants ◽  
Systemic Symptom ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Transmission Electron ◽  
Staining Methods ◽  
Polymerase Chain ◽  
Systemic Symptoms

Abstract Chrysanthemum is one of the important ornamental plants in worldwide due to its high economic and cultural values. Chrysanthemum leaves showed mosaic, ringspot, yellowing and mild mottle symptoms were observed and collected from cultivation areas in northern Thailand and used for detection of important viruses infecting chrysanthemum. Chrysanthemum virus B (CVB) was detected by reverse transcription polymerase chain reaction (RT-PCR) from samples showing yellowing and mild mottle symptoms. Sequences of the coat protein (CP) gene of two CVB isolates found in this study were sequenced and shared 93.15% homology with other CVB isolates from different countries deposited in GenBank. Biological indexing of these CVB found that they induced both local and systemic symptoms in tobacco plants while petunia displayed a systemic symptom. The particles of CVB were observed under transmission electron microscope (TEM), prepared by dip preparation and negative staining methods, showing slightly flexuous rod-shaped virions approximately 600–650 nm in length. To our knowledge, this is the first detection and study on molecular and biological characteristics of CVB infecting chrysanthemum in Thailand.

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.

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