High-resolution melt-curve analysis of random-amplified-polymorphic-DNA markers, for the characterisation of pathogenic Leptospira.

A new test for pathogenic Leptospira isolates, based on RAPD-PCR and high-resolution melt (HRM) analysis (which measures the melting temperature of amplicons in real time, using a fluorescent DNA-binding dye), has recently been developed. A characteristic profile of the amplicons can be used to define serovars or detect genotypes. Ten serovars, of leptospires from the species Leptospira interrogans (serovars Australis, Robinsoni, Hardjo, Pomona, Zanoni, Copenhageni and Szwajizak), L. borgpetersenii (serovar Arborea), L. kirschneri (serovar Cynopteri) and L. weilii (serovar Celledoni), were typed against 13 previously published RAPD primers, using a real-time cycler (the Corbett Life Science RotorGene 6000) and the optimised reagents from a commercial kit (Quantace SensiMix). RAPD-HRM at specific temperatures generated defining amplicon melt profiles for each of the tested serovars. These profiles were evaluated as difference-curve graphs generated using the RotorGene software package, with a cut-off of at least 8 'U' (plus or minus). The results demonstrated that RAPD-HRM can be used to measure serovar diversity and establish identity, with a high degree of stability. The characterisation of Leptospira serotypes using a DNA-based methodology is now possible. As an objective and relatively inexpensive and rapid method of serovar identification, at least for cultured isolates, RAPD-HRM assays show convincing potentia.

The biology of Australian weeds, Limnocharis flava

The genus name Limnocharis is derived from the Greek limno (meaning marsh or pond) and charis (meaning grace) (Haynes and Holm-Nielson 1992) and flava is Latin for yellow. The genus is generally accepted to have two species, Limnocharis flava (Linneaus) Buchenau 1868 and L. laforestii (Duchass. ex Griseb) 1858. L. flava was first named Alisma flava by Linneaus in 1753 (Haynes and Holm-Nielsen 1986). Since then, other synonyms have included Damasonium flavum Mill. 1772, Limnocharis emarginata Humb. and Bonpl. 1808, Limnocharis plumieri Rich. 1815, Limnocharis laforestii Duchas. ex Griseb (1858) and Limnocharis mattogrossensis O. Ktze. (1893) (Woodson and Schery 1943).