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.

Great Barrier Reef Fish Biological Data

Biological measurements on fish sampled during the course of FRDC funded project Growth, Reproduction and Recruitment of Great Barrier Reef Food Fish Stocks (FRDC 90/18). The comma-delimited ascii file comprises the following fields: 1. Cruise number 2. Date (d-m-y) 3, Region (descriptor of part of Queensland coast or Great Barrier Reef system) 4. Reef (name or number) 5. Data source (Res=research, Rec=recreational fisher, Com=commercial fisher) 6. Capture method 7. Trap number (where appropriate) 8. Species name 9. LthStd (standard length, cm) 10. LthFrk (fork length, cm) 11. LthTot (total length, cm) 12. WtTot (approx total weight, g; weighed at sea) 13. FrameWt (weight of frame [after filleting, with viscera], g; weighed in lab) 14. Sex (macroscopic examination only) 15. GonadWt (g) Data obtained by the Department Employment, Economic Development and Innovation (formerly Primary Industries and Fisheries) between 1988 and 1993, primarily in the southern Great Barrier Reef (Capricorn-Bunker and Swain Groups), with fish traps and handlining.