Investigating individual variations between different isolates of group A streptococci, we observed a close correlation between biotypes and serotypes in 46 strains from pharyngitis patients. Biotyping, carried out with a commercially available rapid identification gallery, delineated 10 different associations of characteristics, designated biotypes 1 to 10, observed both in the manufacturer's (127 strains) and our personal (98 strains) collections of group A strains. Only the most frequent biotypes (biotypes 1 to 6) were observed in the pharyngitis cohort, but the overall frequencies of the biotypes did not display striking differences compared with the control collections. Serotyping of the pharyngitis strains showed that each M type was restricted to a sole biotype. For example, M types 1, 4, and 28 were found only in biotype 1 and M type 6 was found only in biotype 6 strains. This association was not due to an epidemiologic bias, since it was also observed in a control series consisting of reference strains and isolates from distant countries (the United States and Czech Republic versus France). An exception was for M type 78, which exhibited biotype 3 or biotype 4. Investigation of the heterogeneity of the strains at the DNA level showed no significant variations of the ribotype patterns between strains of different biotypes, confirming that group A streptococci belong to a unique and homogeneous species. This previously undescribed association between serotypes and biotypes is of interest for a rapid and preliminary characterization of strains isolated in individual patients or during an outbreak. A possible pathogenic association of some biotypic characteristics with specific M proteins is envisaged.Group A streptococci (Streptococcus pyogenes) are the most frequent cause of human pharyngitis (13). This species includes several serotypes which can be specifically involved in acute infections, such as bacteremia and toxic shock-like syndrome, or cause severe complications: nephritis, chorea, and rheumatic fever (12,13,22). Since the 1980s, important changes have arisen in the incidence and clinical features of invasive S. pyogenes infections, and a new emergence of rheumatic fever has been observed all over the world (2,14,17). Recent epidemiological findings have reinforced the interest in this species and led to reassessment of both the efficiency and significance of the methods of strain characterization. The usual classification has been based on antigenic variations of two surface molecules, M protein (15) and T protein (7), and allows delineation between more than 80 and 20 different serotypes, respectively (5); with the presence, or absence, of the serum opacity factor (OF) being correlated with the M protein typing (18).In this study, new typing methods were used to reappraise the homogeneity of the species and to investigate individual variations between different isolates. The S. pyogenes strains are confirmed as belonging to a unique species, with a close correlation between biotypes and serotype...