Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus

Tomás, Gonzalo; Hernández, Martı́n; Marandino, Ana; Techera, Claudia; Grecco, Sofía; Hernández, Diego; Banda, Alejandro; Panzera, Yanina; Pérez, Ruben

doi:10.1080/03079457.2016.1228827

Cited by 10 publications

(7 citation statements)

References 44 publications

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“…Argentine (n = 13) and Uruguayan (n = 11) dIBDV strains collected between 2009 and 2016 were used for partial or complete coding genome amplification (Table 1). Samples were diagnosed and genotyped using previously described RT-qPCR assays (Tomás et al, 2012(Tomás et al, , 2017. For partial genome sequencing, 492-and 594-bp fragments corresponding to the hypervariable region of VP2 (hvVP2) and VP1 genes, respectively, were amplified using previously described primers and conditions (Tomás et al, 2019).…”

Section: Sequencing Of Dibdv Strainsmentioning

confidence: 99%

Origin and global spreading of an ancestral lineage of the infectious bursal disease virus

Tomás

Marandino

Techera

et al. 2019

Transbound Emerg Dis

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Infectious bursal disease virus (IBDV) is an economically relevant and widespreadpathogen that produces immunosuppression in young chickens. IBDV is genetically classified into seven genogroups (G1-G7), where the traditional classic, variant and very virulent strains correspond to G1, G2 and G3, respectively. The G4 strains, also known as 'distinct' (dIBDV), have recently acquired increased relevance because of their prevalence and notorious impair to the poultry industry in South America. Here, worldwide dIBDV strains were studied using phylogenetic and phylodynamic approaches. The phylogenetic analyses performed using partial and complete sequences of both viral segments (A and B) consistently clustered the dIBDV strains in a monophyletic group. The analyses of the VP5, polyprotein and VP1 coding regions identified amino acid residues that act as markers for the identification of the entire dIBDV group or different sub-populations. The phylodynamic analyses performed using the hypervariable region of VP2 indicated that the dIBDV strains emerged in the early 1930s in Eastern Europe, shortly after the emergence of classic strains (1927) and before variant (1949) and very virulent strains (1967). The analysis of the migration routes indicated that after its emergence, the dIBDV strains spread to Eastern Asia around 1959, to Brazil around 1963, and to Argentina around 1990. These inter-continental migrations resulted in three sub-populations that are currently represented by strains from (a) Brazil, (b) Eastern Asia and Canada, and (c) Eastern Europe, Argentina and Uruguay. Taken together, our results highlight the complex evolutionary history of IBDV and the importance of new phylodynamic data to unravel and nearly follow the different evolutionary pathways taken by this important poultry pathogen.

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Section: Sequencing Of Dibdv Strainsmentioning

confidence: 99%

Origin and global spreading of an ancestral lineage of the infectious bursal disease virus

Tomás

Marandino

Techera

et al. 2019

Transbound Emerg Dis

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“…The DNA was stored at -20 °C and later used as the template for sequencing. Extracted FAdVs DNA was tested by PCR and RT-PCR to confirm the absence of other pathogens such as avian reovirus (ARV) [39], chicken anemia virus (CAV) [2], infectious bursal disease virus (IBDV) [38], Marek's disease (MD) [28], and plaque passage was performed.…”

Section: Calculation Of Tcid 50mentioning

confidence: 99%

A comparative pathogenicity analysis of two adenovirus strains, 1/A and 8a/E, isolated from poultry in Poland

Niczyporuk

Czekaj

2018

Arch Virol

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Fowl adenoviruses (FAdVs) are the causative agents of multietiological syndromes and diseases in poultry flocks. During a routine diagnostic examination, two FAdVs strains were isolated. Molecular typing of these isolates based on the partial loop L1 HVR1-4 region of the hexon gene sequence revealed the presence of different FAdV isolates: 1/A-61/11z (GenBank accession number KX247012, APP94082), and 8a/E-6/12j (GenBank accession number: KP890032, ALB00550), and comparative genome analysis indicated small differences between these two viruses. The next step of the study was the estimation of the pathogenicity of these isolates in specific-pathogen-free (SPF) chickens. Chickens were divided into three groups, with 20 chickens per group infected intraperitoneally on the first day after hatching. Group I consisted of chickens infected with strain FAdV-1/A-61/11z, group II consisted of chickens infected with strain FAdV-8a/E-6/12j, and group III consisted of uninfected birds. Clinical signs observed in infected chickens included poor growth, apathy, prostration, ruffled feathers, crouching position, and huddling behavior. The mortality rate in chickens infected with FAdV-1/A-61/11z was 10% at 10 days postinfection (dpi), and no mortality was observed in chickens infected with the FAdV-8a-6/12j strain. The mean real-time PCR threshold cycle (Ct) value was 39.70%. The detection limit of these assays was 8 copies, with an efficiency of 91.03% and 95.17% and regression square (R2) values of 0.991 and 0.997, respectively, with a mean pathogen load of 4.8 × 10 6.0 copies/µl. The assays did not demonstrate cross-reactivity between types 1/A and 8a/E and non-targeted poultry viruses. Adenoviral DNA was detected in the liver, spleen, kidney, gizzard, intestine, bursa of Fabricius, and thymus of every examined dead and euthanized chicken from groups I and II between the third and fourth week postinfection. This is the first study conducted on the pathogenic and apathogenic strains FAdV-1/A and FAdV-8a/E, showing the presence of the virus in multiple tissues in chickens in Poland. This study revealed that it is very likely that the FAdV-1/A-61/11z strain is able to cause clinical inclusion body hepatitis (IBH) in chickens and that it is slightly more virulent than the FAdV-8a/E-6/12j strain, although both are primary pathogens of the disease.

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“…The application of rRT-PCR as a tool for the diagnosis of IBDV infections has been used [14] because of its simplicity, high sensitivity and specificity [15]. The assay is usually carried out with the use of hydrolysis probe (Taq-Man) [16].…”

Section: Diagnosis Of Clinical Cases Of Infectious Bursal Disease Usimentioning

confidence: 99%

Diagnosis of Clinical Cases of Infectious Bursal Disease Using a Modified Rapid Taq Man-MGB Real-Time RT-PCR Assay

Cheggag¹,

Zro²,

Sebbar³

et al. 2018

JAST-A

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Infectious bursal disease (IBD) is an important contagious viral infection of immune system of poultry. This infection possesses a permanent threat to the profitability of poultry industry worldwide. The aim of this work was to modify the Taq Man-MGB real-time reverse transcription-polymerase chain reaction (rRT-PCR) in one step involving two fluorogenic Taq Man labeled probe and using this protocol for detection of infectious bursal disease virus (IBDV) collected from suspected cases distributed in different regions of the country during the period 2013-2016. The intralaboratory validation of modified method was realized for specificity, linearity, repeatability, sensitivity and reproducibility. It allowed reducing the test running time by six folds. This method was applied on 102 pools of bursa of fabricius (BF) samples collected from affected broiler farms suspected to be infected by IBDV. Birds showing macroscopic lesions including muscle petechial hemorrhages, hypertrophy and hemorrhage of BF, were subjected to molecular analysis using modified protocol "Taq Man-MGB rRT-PCR". The validation satisfied all criteria and the assay developed could be a useful tool for a very rapid diagnosis of IBDV and permit to detect and to discriminate in one-step very virulent (vv) from non-vv (classic and variant) IBDV strains. Out of 84 IBDV positive samples, a prevalence of 39% for vv strains and 61% for classical strains was noted. These results indicate that despite the vaccination against IBDV, the vv form of this pathologie continues to cause serious problems for Moroccan broiler chickens. The obtained results indicate the successfully detection of IBDV and differentiated all vvIBDV strains from non-vvIBDV strains; Avian infectious agent RNA viruses tested are negative, demonstrating great specificity of the assay. The results obtained indicate that this method is suitable as a routine laboratory test for the rapid detection and differentiation of IBDV strains in samples of avian origin.

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Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus

Cited by 10 publications

References 44 publications

Origin and global spreading of an ancestral lineage of the infectious bursal disease virus

Origin and global spreading of an ancestral lineage of the infectious bursal disease virus

A comparative pathogenicity analysis of two adenovirus strains, 1/A and 8a/E, isolated from poultry in Poland

Diagnosis of Clinical Cases of Infectious Bursal Disease Using a Modified Rapid Taq Man-MGB Real-Time RT-PCR Assay

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