Simultaneous detection of astrovirus, rotavirus, reovirus and adenovirus type I in broiler chicken flocks

2012 ◽  
Vol 15 (2) ◽  
pp. 337-344 ◽  
Author(s):  
D. Roussan ◽  
I. Shaheen ◽  
G. Khawaldeh ◽  
W. Totanji ◽  
R. Al-Rifai
Keyword(s):  
Polymerase Chain Reaction ◽  
Broiler Chicken ◽  
Simultaneous Detection ◽  
Adenovirus Type ◽  
Economic Losses ◽  
Type I ◽  
Chain Reaction ◽  
Group I ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Simultaneous detection of astrovirus, rotavirus, reovirus and adenovirus type I in broiler chicken flocksEnteric diseases cause substantial economic losses to the poultry industry. Astroviruses, rotaviruses, reoviruses, and adenovirus type 1 have been reported as a significant cause of intestinal symptoms in poultry. In the present study, intestinal samples from 70 commercial broiler chicken flocks were examined for the presence of astroviruses, rotavirus, and reovirus by reverse transcription-polymerase chain reaction, and for the presence of group I adenovirus by polymerase chain reaction. Astroviruses were identified in 38.6% of samples tested. Both avian nephritis virus and chicken astrovirus were identified in the astrovirus positive flocks, where 74.1% of these flocks were positive for only one type of astrovirus, whereas, 25.9% of these flocks were positive for both types of astrovirus. Reoviruses, rotaviruses, and adenoviruses were identified in 21.4, 18.6, and 14.3% of these flocks, respectively. Concomitant infection with two or more viruses in the same flock were also prominent, where 5.7, 5.7, 2.9, 2.9, 1.4, and 1.4% of these flocks were positive with both astrovirus and rotavirus; astrovirus and adenovirus; astrovirus and reovirus; rotavirus and adenovirus; rotavirus and reovirus; and reovirus and adenovirus respectively. Moreover, 4.3 and 2.7% of these flocks were positive for astrovirus, reovirus, and adenovirus; and astrovirus, reovirus, and rotavirus, respectively. Further studies will focus on identifying specific viral factors or subtypes/subgroups associated with disease through pathogenesis studies, economic losses caused by infections and co-infections of these pathogens, and the costs and benefits of countermeasures.

scholarly journals Sensitive SYBR Green—Real Time PCR for the Detection and Quantitation of Avian Rotavirus A

2018 ◽  
Vol 6 (1) ◽  
pp. 2 ◽  
Author(s):  
David De la Torre ◽  
Claudete Astolfi-Ferreira ◽  
Ruy Chacon ◽  
Antonio Piantino Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Sybr Green ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rotavirus A ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Avian rotavirus A (ARtV-A) is a virus that affects young birds, causing acute diarrhea and economic losses in the poultry industry worldwide. The techniques used for the diagnosis of ARtV-A include electron microscopy, isolation in cell culture, and serology, as well as molecular techniques, such as the reverse transcription-polymerase chain reaction (RT-PCR). The objective of this work was to standardize a real-time RT-polymerase chain reaction (RT-qPCR) using SYBR Green chemistry for the rapid detection and quantification of ARtV-A from bird tissues and materials fixed on FTA cards on the basis of the nucleotide sequence of segment 6 (S6), which codes for the structural VP6 protein of ARtV-A. The results show the efficient amplification of the proposed target, with a limit of detection (LoD) of one copy gene (CG) per microliter of cDNA and a limit of quantification (LoQ) of 10 CGs per microliter. The efficiency of the primers was determined to be 95.66% using a standard curve, with an R2 value of 0.999 and a slope of −3.43. The specificity was determined using samples coinfected with ARtV-A, the chicken parvovirus, the chicken astrovirus, and the avian nephritis virus as positive controls and commercially available vaccines of the infectious bronchitis virus, infectious bursa disease virus, avian reovirus and healthy organs as negative controls. This technique, which lacks nonspecific PCR products and dimers, demonstrated greater sensitivity and specificity than conventional RT-PCR, and it reduced the analysis time by more than 50%.

scholarly journals Comparison of polymerase chain reaction and microscopy for the detection of Fasciola spp. in the fecal matter of domestic bovines in Kalasin Province, Thailand

2021 ◽  
pp. 2878-2882
Author(s):  
Sirikanda Thanasuwan ◽  
Anupong Tankrathok
Keyword(s):  
Polymerase Chain Reaction ◽  
False Negative ◽  
Economic Losses ◽  
Rrna Gene ◽  
28S Rrna ◽  
Chain Reaction ◽  
Fecal Matter ◽  
Specificity And Sensitivity ◽  
Polymerase Chain ◽  
Fasciola Spp

Background and Aim: Fasciola spp. are important foodborne trematodes and waterborne zoonotic parasites that cause health problems and economic losses worldwide, including in Thailand. Fasciola spp. are usually detected by sedimentation or the formalin-ethyl acetate concentration technique (FECT) under microscopy, which is less specific and sensitive. Accurate detection is important to detect real incidence for protection against and elimination of fasciolosis in the area. This study aimed to determine the distribution of Fasciola spp. and compare the specificity and sensitivity of FECT under microscopy to that of polymerase chain reaction (PCR) in cattle feces. Materials and Methods: The study was conducted in Kalasin Province, Thailand. Feces of 46 cattle were investigated for infection with Fasciola spp. To detect infection, FECT under microscopy and PCR amplification of the 28S rRNA gene of Fasciola spp. were used to identify egg parasites. Results: Feces of 16 of 46 (34.78%) cattle were positive for Fasciola spp. using FECT under microscopy, whereas PCR showed that 67.39% (31 of 46) were positive for Fasciola spp. False-negative results were as high as 32.61% when diagnosed under microscopy. Conclusion: This study confirmed the infection of cattle with Fasciola spp. in Kalasin Province, indicating that PCR demonstrated higher sensitivity and specificity when diagnosing infection. FECT under microscopy can still be used as a primary and traditional method for diagnosis. However, relapse cases of Fasciola spp. and Paramphistomum spp. should be diagnosed by microscopy combined with PCR. This is the first report on the molecular distribution of fecal samples in cattle in Kalasin Province.

scholarly journals Histopathological and virological findings in emaciated pigs from Mexico: an exploratory study

2018 ◽  
Vol 87 (3) ◽  
pp. 213-217
Author(s):  
Aide Alpízar ◽  
Joaquim Segalés ◽  
Simón Martínez ◽  
Atalo Martínez ◽  
Guadalupe Socci ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Body Condition ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Economic Losses ◽  
Respiratory Syndrome Virus ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Histopathological Studies ◽  
Weaning Piglets

The objective of this work was to detect the presence of three main pig respiratory viral agents (porcine rubulavirus [PorPV], porcine circovirus type 2 [PCV-2], and porcine reproductive and respiratory syndrome virus [PRRSV]) in tissues of emaciated piglets from the Baj'o Region (Mexico). Necropsies and histopathological studies of 37 pigs with poor body condition were performed; viruses were detected by molecular biology methods and PCV-2 was further assessed by immunohistochemistry (IHC). Histopathologically, interstitial pneumonia was observed in 25/37 (68%) of the piglets. Also, a varying degree of lymphocyte depletion in lymphoid organs was found in 14/37 (38%) animals. Through polymerase chain reaction (PCR) and/or reverse transcription polymerase chain reaction (RT-PCR), from the 37 pigs, 16 were positive for PCV-2, 18 for PRRSV and 1 for PorPV. In accordance with these results, the infection and/or co-infection with PCV-2 and PRRSV were fairly frequent findings in piglets with poor body condition in Mexico, while the infection by PorPV was apparently negligible. Wasting of post-weaning piglets is a global pig farming problem that causes great economic losses and has been associated with diverse factors: microbial agents, environmental factors, nutritional factors, and management. When the Blue Eye Disease was first reported in Mexico, it was associated with severe wasting in post-weaning piglets. This study demonstrated that this disease does not seem to play such an important role in the wasting as was previously thought.

scholarly journals Quantification of Venturia inaequalis Growth in Malus × domestica with Quantitative Real-Time Polymerase Chain Reaction

Plant Disease ◽  
2012 ◽  
Vol 96 (12) ◽  
pp. 1791-1797 ◽  
Author(s):  
Michele Gusberti ◽  
Andrea Patocchi ◽  
Cesare Gessler ◽  
Giovanni A. L. Broggini
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Malus Domestica ◽  
Venturia Inaequalis ◽  
Simultaneous Detection ◽  
Scab Resistance ◽  
Infected Leaf ◽  
Chain Reaction ◽  
Limit Of Quantification ◽  
Polymerase Chain

A quantitative real-time polymerase chain reaction (qPCR) was developed and validated for quantification of Venturia inaequalis in infected leaf tissue of Malus × domestica. The method is based on dual-labeled hybridization probes, allowing simultaneous detection of host and pathogen DNA within one single reaction. Limit of quantification for the pathogen was 0.5 pg per reaction and, for the host, reached 5 pg per reaction. The fungal growth measured in four apple cultivars 2 weeks after inoculation significantly correlated with their different level of scab resistance and allowed the observation of ontogenic resistance. After sporulation on the youngest leaf, fungal biomass in susceptible ‘Gala’ was 118 times higher than in resistant ‘Florina’ and ‘Discovery’ while intermediate values were found with the intermediate susceptible ‘Milwa’. Correlation was also observed between severity classes obtained by visual scoring of symptoms and qPCR results. Moreover, qPCR demonstrated validity of the developed method as a disease severity forecast tool 10 days after the pathogen's inoculation and prior to the appearance of the symptoms. Applications of the methodology can include the quantification of scab resistance during breeding programs, evaluation of fungicide and biocontrol efficacy, and quantification of the fitness of different pathogenic strains.

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