New polymerase chain reaction (PCR) primers were developed for the identification of the EU quarantine pest Monilinia fructicola. These allowed nine M. ffucticola isolates to be distinguished from other fungi, including six isolates of M. laxa and six isolates of M. fructigena (which also cause brown rot of stone and pome fruit). Three M. fnrcticola isolates from Japan and one from Australia did not react with a primer set previously published for M. fructicola. M. fnrcticola isolates could be subdivided into three groups based on the size of their nuclear rDNA small subunit. The subunits from the four non-reactive isolates were either smaller or larger than the reactive group. Further primers were developed which were specific for either M. laxa or M. fructigena. Another new primer identified both M. laxa and M. fructigena, and yet another M. laxa and M. fructicola. When used in combination, these primers specific for two species correctly identified unknown isolates of all three Monilinia species. The new primers designed in this study have been used to identify, rapidly and correctly, pustules taken directly from infected plum fruits, thus demonstrating their diagnostic potential. and 2 regions of the nuclear rRNA gene repeat were therefore produced ( Table 1). The effectiveness of these primers and the occurrence of group-I introns in isolates of M. fructicola is discussed.

Monilinia fructicola, M. laxa and M. fructigena are the causal agents of brown rot of pome and stone fruits. M. fructicola is not present in Europe and is classed as a quarantine pathogen in EU countries. A 418-bp group-I intron has been located in the small subunit (SSU) rDNA gene of M. fructicola which is absent from M. laxa and M. fructigena. PCR primers specific to the 3'-region of the intron together with the SSU rDNA primer NS5 were able to amplify a 444-bp product from M. fructicola and fruit tissue infected with M. fructicola but not from the other two species. This allows for the rapid and sensitive detection of this pathogen in planta.

Background: Non-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. At present no reliable biomarkers are available to guide the management of this condition. Microarray technology may allow appropriate biomarkers to be identified but present platforms are lacking disease focus and are thus likely to miss potentially vital information contained in patient tissue samples.

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PCR amplification of the small subunit (SSU) rDNA gene of 40 isolates of Nectria galligena revealed four length polymorphisms. PCR-RFLP analysis of the SSU rDNA gene divided the isolates into four categories similar, but not identical, to categories identified by Southem-RFLP analysis. Nucleotide sequence analysis revealed that isolates in three of the four SSU rDNA (18s) categories possess an intron of 363 bp, 1185 bp or 1423 bp at the NS 7 priming site. Isolates in the fourth category do not possess an intron. The nucleotide sequences of these introns did not contain the core elements characteristic of typical group I introns, nor did they exhibit a group I intron secondary structure. Homology between the introns indicates a common lineage, all three possibly having come from a larger intron and having been formed by subsequent deletions. PCR primers upstream of the SSU rDNA intron region and from within the internal transcribed spacer 1 region amplify a product specific to N. ga//igena, which will confirm the identity of the pathogen and reveal its 185 category in a single reaction.