title: Application of sensitive and specific molecular methods to uncover global dissemination of the major RDRio Sublineage of the Latin American-Mediterranean Mycobacterium tuberculosis spoligotype family.
abstract: The Latin American-Mediterranean (LAM) family of Mycobacterium tuberculosis is believed to be the cause of approximately 15% of tuberculosis cases worldwide. Previously, we defined a prevalent sublineage of the LAM family in Brazil by a single characteristic genomic deletion designated RD(Rio). Using the Brazilian strains, we pinpoint an Ag85C(103) single nucleotide polymorphism (SNP) (screened by restriction fragment length polymorphism [RFLP] analysis) that correctly identified all LAM family strains. Importantly, all RD(Rio) strains concomitantly possessed the RD174 deletion. These genetic signatures, along with a newly developed multiplex PCR for rapid differentiation between "wild-type" and RD(Rio) strains, were then used to analyze an international collection of M. tuberculosis strains. RD(Rio) M. tuberculosis was identified from four continents involving 11 countries. Phylogenetic analysis of the IS6110-RFLP patterns from representative RD(Rio) and LAM strains from Brazil, along with all representative clusters from a South African database, confirmed their genetic relatedness and transcontinental transmission. The Ag85C(103) SNP RFLP, as compared to results obtained using a PCR method targeting a LAM-restricted IS6110 element, correctly identified 99.8% of LAM spoligotype strains. Together, these tests were more accurate than spoligotyping at categorizing strains with indefinable spoligotypes and segregated true LAM strains from those with convergent spoligotypes. The fact that RD(Rio) strains were identified worldwide highlights the importance of this LAM family sublineage and suggests that this strain is a global threat that should be specifically targeted by public health resources. Our provision of simple and robust molecular methods will assist the evaluation of the LAM family and the RD(Rio) sublineage.
authors.affiliation: Cornell University, Joan and Sanford I. Weill Medical College, Department of Medicine, Division of International Medicine and Infectious Diseases, Rm. A-421, 525 East 68th St., New York, NY 10065, USA. jlho@med.cornell.edu
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Gibson
authors.forename: Richard C
authors.initials: RC
authors.lastname: Huard
authors.forename: Nicolaas C
authors.initials: NC
authors.lastname: Gey van Pittius
authors.forename: Luiz Claudio Oliveira
authors.initials: LC
authors.lastname: Lazzarini
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Driscoll
authors.forename: Natalia
authors.initials: N
authors.lastname: Kurepina
authors.forename: Thierry
authors.initials: T
authors.lastname: Zozio
authors.forename: Christophe
authors.initials: C
authors.lastname: Sola
authors.forename: Silvana Miranda
authors.initials: SM
authors.lastname: Spindola
authors.forename: Afrânio L
authors.initials: AL
authors.lastname: Kritski
authors.forename: Daniel
authors.initials: D
authors.lastname: Fitzgerald
authors.forename: Kristin
authors.initials: K
authors.lastname: Kremer
authors.forename: Helmi
authors.initials: H
authors.lastname: Mardassi
authors.forename: Poonam
authors.initials: P
authors.lastname: Chitale
authors.forename: Jessica
authors.initials: J
authors.lastname: Brinkworth
authors.forename: Dario
authors.initials: D
authors.lastname: Garcia de Viedma
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gicquel
authors.forename: Jean W
authors.initials: JW
authors.lastname: Pape
authors.forename: Dick
authors.initials: D
authors.lastname: van Soolingen
authors.forename: Barry N
authors.initials: BN
authors.lastname: Kreiswirth
authors.forename: Robin M
authors.initials: RM
authors.lastname: Warren
authors.forename: Paul D
authors.initials: PD
authors.lastname: van Helden
authors.forename: Nalin
authors.initials: N
authors.lastname: Rastogi
authors.forename: Philip N
authors.initials: PN
authors.lastname: Suffys
authors.forename: Jose
authors.initials: J
authors.lastname: Lapa e Silva
authors.forename: John L
authors.initials: JL
authors.lastname: Ho
mesh_terms: D000217:Acyltransferases / Q000235:genetics; D000942:Antigens, Bacterial / Q000235:genetics; D003062:Codon; D004251:DNA Transposable Elements / Q000235:genetics; D019143:Evolution, Molecular; D064429:Fatty Acid Synthases / Q000235:genetics; D014943:Global Health; D006801:Humans; D007843:Latin America; D019083:Mediterranean Region; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D009841:Oligonucleotides / Q000032:analysis*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000453:epidemiology* / Q000382:microbiology / Q000635:transmission*
chemical_list: D000942:Antigens, Bacterial; D003062:Codon; D004251:DNA Transposable Elements; D009841:Oligonucleotides; D000217:Acyltransferases; C119774:antigen 85C, Mycobacterium tuberculosis; D064429:Fatty Acid Synthases