Authors:J. Kundu, J. Jarošová, S. Gadiou, and G. Cervená
One-step RT-PCR followed by a RFLP assay was developed for the typing of
Barley yellow dwarf virus
(BYDV) member of
genera. RFLP analysis performed on 23 samples including one lab isolate showed three types of
II restriction profile. A partial coat protein (CP) gene sequencing was carried out and confirmed the RFLP analysis. Both sequencing and RFLP analysis identified the presence of 3 BYDV-PAV (including the lab isolate Blatno85), 10 BYDV-MAV and 10 BYDV-PAS isolates among the field samples. One-step RT-PCR together with RFLP presents an easy and reliable assay for routine BYDV typing. These methods revealed that PAS and MAV are more dominant than PAV in the Czech Republic.
Authors:J. Krulj, N. Ćurčıć, A. Bočarov Stančıć, J. Kojıć, L. Pezo, L. Peıć Tukuljac, and M. Bodroža Solarov
sequences for the description and identiﬁcation of the species using a multilocus approach. PCR-restriction fragment length polymorphism (PCR-RFLP) is a simple, cost eﬀective and quick tool for rapid detection of speciﬁc diﬀerences in DNA sequences of
Authors:Boglárka Sellyei, Éva Ivanics, and Tibor Magyar
The 16 somatic serotype type strains and 60 field isolates of Pasteurella multocida, representing various avian species and geographic regions in Hungary, were characterised by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the ompH gene with DraI restriction endonuclease. The type strains yielded eight different (I-VIII) profiles. Strains whose PCR fragment was uncut by DraI (profile IV) could be differentiated with HindIII and PvuII restriction endonucleases. Five of the eight PCR-RFLP profiles (I, III, V, VI and VII) were detected among the field strains. Only a correlation of limited strength was found between the classical somatic serotypes and the PCR-RFLP profiles. However, the results confirmed that molecular methods could confidently distinguish serotype A:1 strains from the other serotypes. Moreover, the specific relationship between somatic serotypes and PCR-RFLP types among isolates from turkey raises the possibility of the existence of host-specific clones within the P. multocida population.
Authors:Ángela Lacombe-Antoneli, Segundo Píriz, Alberto Quesada, and Santiago Vadillo
Footrot is widely considered the most severe and most common foot pathology in small ruminants. This study tested the ability of a molecular typing system based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay of the 16S rRNA gene to discriminate between the strict anaerobe genera most commonly isolated from footrot (
Bacteroides, Dichelobacter, Fusobacterium, Porphyromonas
) in goats in Extremadura (Spain), with a view to facilitating identification for diagnostic purposes and thus providing a useful tool for future epidemiological studies. Although the electrophoretic patterns obtained with the enzyme
1I were more readily interpreted, and may thus be the best initial option, results may be confirmed by a second enzyme (
I). The PCR-RFLP assay of the 16S rRNA gene may therefore prove a useful addition to conventional biochemical identification techniques, providing taxonomic information at genus level.
Authors:Zsuzsanna Tapaszti, Petra Forgách, Csaba Kővágó, László Békési, Tamás Bakonyi, and Miklós Rusvai
Microsporidiosis (nosema disease) of the European honeybee (
L.) is present in bee colonies worldwide. Until recently,
had been regarded as the causative agent of the disease, which may have many negative effects on the colony and cause heavy economic losses in apicultures. Another microsporidium species,
, was reported to infest the Asian honeybee (
), but both honeybee species are susceptible to both microsporidia. In the European honeybee
was first detected in Spain in the year 2006. As it is difficult to distinguish
morphologically, a rapid and accurate assay has been developed to differentiate
based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the partial large subunit ribosomal RNA. The assay was tested on 38
-infested bee samples, which were collected from geographically distant Hungarian bee colonies representing all regions of the country. Only one sample contained
, and in the other 37 samples
was detected, which indicates the dominance of
in Hungarian apiaries. This is the first report on the presence of
Jahoor, A., Jacobi, A., Schüller, C. M. E., Fischbeck, G. 1993: Genetical and RFLP studies at the Mla locus conferring powdery mildew resistance in barley. Theor. Appl. Genet. , 85 , 713-718.
Genetical and RFLP studies at the
Wine fermentation is a complex microbiological process in which yeasts predominate. It is long debated whether yeasts occurring on the surface of grapes or the resident yeasts on the winery equipment play the primary role in conducting the fermentation. The origin, development, changes and succession of various yeast species can be followed using specific molecular techniques allowing the differentiation and typing of yeast strains. Techniques such as pulsed field gel electrophoresis of chromosomal DNA, restriction fragment length polymorphism analysis, and polymerase chain reaction (PCR)-based methods have recently been employed in studying the microbiology of wine making. These shed new light on the dynamics of fermentation started spontaneously or directed by the inoculation of starter cultures.