Typing of Haemophilus paragallinarum Strains by Using Enterobacterial Repetitive Intergenic Consensus-Based Polymerase Chain Reaction

This study examined 49 field isolates of the genus Avibacterium, with the 49 being allocated to 36 epidemiologically unrelated groups and one isolate from each group being examined in detail. In addition, six type and reference strains were investigated. Phylogenetic analysis of partially sequenced recN, rpoB, infB, pgi and sodA genes confirmed the existence of the species Avibacterium paragallinarum, while a species complex encompassing Avibacterium volantium, Avibacterium avium, Avibacterium gallinarum, Avibacterium endocarditis and Avibacterium sp. A could not be resolved. All isolates shared at least one identical sequence in one gene, indicating low diversity or horizontal gene transfer (HGT) between isolates. Such HGT between isolates of defined species and unclassified isolates combined with high sequence similarity can be explained as the result of an ongoing speciation process. The alternative explanation is that Av. volantium, Av. avium and Avibacterium sp. A were misclassified originally. Except for Av. paragallinarum, identification of species of Avibacterium seems problematic, even by DNA sequencing, as shown in the present investigation. The results indicate that Avibacterium probably contains only two or three species. Until the taxonomic revision is completed we recommend that isolates that do not fit with named species by genotype and phenotype be designated Avibacterium sp.

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“…Enterobacterial repetitive intergenic consensus (ERIC)-PCR of Dutch isolates was performed according to the method of Soriano et al (2004).…”

Section: Methodsmentioning

confidence: 99%

Multilocus sequence phylogenetic analysis of Avibacterium

Bisgaard

Nørskov‐Lauritsen

Wit³

et al. 2012

Microbiology

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“…However, the PCR could not identify serovars A, B and C. Soriano et al reported that an enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) could identify each serovar [13], but it would not be easy to analyze the results, since ERIC-PCR showed many patterns even within each serovar. Therefore, we examined more simplified methods using PCR.…”

mentioning

confidence: 99%

Development of a Multiplex PCR and PCR-RFLP Method for Serotyping of Avibacterium paragallinarum

Sakamoto

Kino

Sakaguchi

2012

The Journal of Veterinary Medical Science

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ABSTRACT. Avibacterium (Haemophilus) paragallinarum (A. paragallinarum) is a causative agent of infectious coryza in chickens and is classified into three serovars by agglutination tests. In an effort to identify the serovars easily, PCR and PCR-RFLP were employed. As the target gene for PCR, the hypervariable region of HMTp210, which encodes the HA antigen, was used. PCR using primer sets around the hypervariable region amplified 0.8, 1.1 and 1.6 kbp fragments for serovars A, B and C, respectively. Alternatively, the 1.6 kbp fragments were amplified with another primer pair encompassing the hypervariable region and was subjected to digestion with Bgl II, which resulted in the detection of serovar-specific digestion patterns. These results indicate that PCR and PCR-RFLP using the hypervariable region of HMTp210 are alternative methods to identify the serovar of A. paragallinarum.

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“…The ERIC-PCR was performed as described by Soriano et al (2004c) using primer pair ERIC1R (5 ′ -ATG-TAAGCTCCTGGGGATTCAC-3 ′ ) as reverse primer and ERIC2 (5 ′ -AAGTAAGTGACTGGGGTGAGCG-3 ′ ) as forward primer. All reactions were performed in duplicate using the same PCR kit and thermocycler as described previously.…”

Section: Enterobacterial Repetitive Intergenic Consensus (Eric)-pcrmentioning

confidence: 99%

“…paragallinarum from other species and does not differentiate serovars within the species (Chen et al, 1996). Molecular typing methods have been reported for the differentiation of different serovars and these methods include Enterobacterial Repetitive Intergenic Consensus (ERIC) polymerase chain reaction (PCR) (Soriano et al, 2004c), Multiplex PCR and Restriction Fragment Length Polymorphism -PCR (Sakamoto et al, 2011). In 2014, research suggested that the Multiplex PCR method is flawed when applied to the identification of unknown serovars of Av.…”

Section: Introductionmentioning

confidence: 99%

“…These different band sizes result in unique banding profiles (Versalovic et al, 1991). During a study by Soriano et al (2004c), the ERIC-PCR was performed on Av. paragallinarum and was used for the differentiation of unknown serovars by comparing banding patterns to group isolates, thus subtyping these strains based on the obtained banding pattern to possibly aid in epidemiological studies and ultimately contribute to disease control.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of the ERIC-PCR as a probable method to differentiate Avibacterium paragallinarum serovars

et al. 2017

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Infectious coryza, an upper respiratory tract disease in chickens, caused by Avibacterium paragallinarum, leads to huge economic losses. The disease is controlled through vaccination; but vaccination efficacy is dependent on correct identification of the infecting serovar, as limited cross-protection is reported amongst some serovars. Current identification methods include the heamagglutination inhibition test, which is demanding and could be subjective. To overcome this, molecular typing methods proposed are the Multiplex polymerase chain reaction (PCR) and Restriction Fragment Length Polymorphism-PCR, but low reproducibility is reported. Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR has been suggested for molecular groupings of various bacterial species. This study focuses on evaluating the ERIC-PCR as a probable method to differentiate between different Av. paragallinarum serovars by grouping with reference isolates, based on clonal relations. The ERIC-PCR was performed on 12 reference isolates and 41 field isolates originating from South Africa and South America. The data indicate that the ERIC-PCR is not ideal for the differentiation or for molecular typing of Av. paragallinarum serovars, as no correlation is drawn upon comparison of banding patterns of field isolates and reference strains. However, the results do indicate isolates from the same origin sharing unique banding patterns, indicating potential clonal relationship; but when compared to the reference isolates dominant in the specific area, no correlation could be drawn. Furthermore, although the ERIC-PCR serves a purpose in epidemiological studies, it has proved to have little application in differentiating amongst serovars of Av. paragallinarum and to group untyped field strains with known reference strains.

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Typing of Haemophilus paragallinarum Strains by Using Enterobacterial Repetitive Intergenic Consensus-Based Polymerase Chain Reaction

Cited by 21 publications

References 12 publications

Multilocus sequence phylogenetic analysis of Avibacterium

Multilocus sequence phylogenetic analysis of Avibacterium

Development of a Multiplex PCR and PCR-RFLP Method for Serotyping of Avibacterium paragallinarum

Evaluation of the ERIC-PCR as a probable method to differentiate Avibacterium paragallinarum serovars

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