Preface

Themycobacteria include a number of important human and animal pathogens and pose major problems worldwide in terms of global health and economies. Tuberculosis poses a significant threat to global health by infecting and killing millions annually. Leprosy has not yet been eradicated, and other infections, such as Buruli ulcer and opportunistic infections associated with immunodeficiency, are on the rise. For these reasons, the need for methods to study the biology of the mycobacteria and to improve diagnostic, therapeutic, and preventative reagents is still a priority. It has been nearly 10 years since the first edition of Mycobacteria Protocols. The response to the first edition was both surprising and pleasing.Many readers commented on how the book had helped them with a tricky problem or allowed them to branch into new areas, and newcomers to the field were able to avoid many common pitfalls and progress quickly. Mycobacteria are difficult organisms to work with, but the availability of the tips and tricks developed by a multitude of scientists over many years has been received very positively. Within my own laboratory, this book has proved invaluable for neophytes. During the time since it was published, research into the mycobacteria has continued to expand, and the number of scientists studying these problematic bacteria has increased. New methods have been developed, and older methods have been refined, making a second edition timely. In this second edition, we have tried to include a range of methods, from the basics of subcellular fractionation, strain typing, and determining minimum inhibitory concentrations (MICs) to more advanced methods using specialized growth conditions and whole genome, transcriptome, and proteome analyses. Although we cannot include all methods used in the modern research laboratory, we hope that this edition will provide a useful primer for new mycobacterial researchers and stimulate new avenues for established scientists. In developing this edition, we have revised and updated some of the most used methods from the original version and incorporated a number of wholly new methods, some of which make use of our expanded horizons in the post– genomic age. Several methods are described forMycobacterium tuberculosis, as this is now the most widely studied species, but can be easily adapted for other members of the genus.