Development of a standardized subgrouping scheme for Legionella pneumophila serogroup 1 using monoclonal antibodies

Abstract

A panel of monoclonal antibodies to Legionella pneumophila serogroup 1 and a subclassification scheme were developed in a collaborative project among three laboratories. The seven most useful monoclonal antibodies were selected from three previously developed panels on the basis. of indirect fluorescent antibody patterns with 83 strains of L. pneumophila serogroup 1 that were obtained from widely distributed geographic locations. The isolates were divided into 10 major subgroups on the basis of reactivity patterns that can be readily reproduced in any laboratory and are not subject to major inconsistencies of interpretation of staining intensity. A standard protocol for the indirect fluorescent antibody procedure was also developed.

Evaluation of an immunomagnetic separation assay in combination with cultivation to improveLegionella pneumophilaserogroup 1 recovery from environmental samples

“…The genomic diversity of these Legionella strains was analysed by arbitrarily primed PCR (AP-PCR) using two primers: 5¢-AAGTAAGTGACTGGG-GTGAGC-3¢ and 5¢-GGTGGTGGCT-3¢, as previously described (Grattard et al 1996). A few strains (see Results) were typed at the CNRL by direct immunofluorescence using MAbs as previously described (Joly et al 1986;Helbig et al 1997).…”

Persistence of chlorine-sensitiveLegionella pneumophilain hyperchlorinated installations

“…Species and serogroup identification of Legionella isolates was performed using immunofluorescence (IF) with rabbit antisera, including 14 serogroups of L. pneumophila and another nine species, as previously described (Pelaz et al 1992). Legionella pneumophila serogroup 1 isolates were subtyped by IF using the International Panel of Monoclonal Antibodies (Joly et al 1986).…”

A multicenter evaluation of genotypic methods for the epidemiologic typing of Legionella pneumophila serogroup 1: results of a pan-European study

“…RFLP I This was performed as described by Gaia et al [21]. Briefly, genomic DNA samples were concomitantly digested with restriction endonucleases PstI and EcoRI (Boehringer) and the digested products separated by electrophoresis on 0.8% agarose (Molecular Biology Certified Agarose; Bio-Rad) gels in 1 XTBE for 16 h at [35][36][37][38][39][40] V. The 100-bp DNA ladder (Life Technologies) was used as molecular size marker. The DNA was then transferred onto a nylon membrane (Biodyne B transfer membrane; Pall Bio-Pharmaceuticals, New York, NYC, USA) and probed with digoxigenin-labeled plasmids.…”

“…The monoclonal subgroups Heysham and Camperdown were not encountered in the study strains using either panel. The different patterns of reactivity of the mAbs in each [38]). Isolates within each of the epidemiologically related sets in subdivisions I and I1 were found to be of the same monoclonal subgroup, i.e.…”