Open AccessSpecific detection of a probiotic Lactobacillus strain in faecal samples by using multiplex PCR
Author(s) -
Lucchini Franco,
Kmet Vladimir,
Cesena Camilla,
Coppi Lucia,
Bottazzi Vittorio,
Morelli Lorenzo
Publication year - 1998
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1998.tb12832.x
Subject(s) - lactobacillus gasseri , biology , lactobacillus , probiotic , microbiology and biotechnology , 16s ribosomal rna , multiplex polymerase chain reaction , primer (cosmetics) , polymerase chain reaction , strain (injury) , bacteria , gene , multiplex , dna extraction , isolation (microbiology) , chemistry , genetics , anatomy , organic chemistry
Lactobacillus gasseri 4B2 is a human isolate characterised by a strong autoaggregating phenotype mediated by APF (aggregation‐promoting factor), a secreted protein. Two primer pairs were developed for simultaneous amplification of a specific fragment of the APF gene and a highly conserved region of the 16S rRNA gene. The specificity of this protocol was checked in DNA samples isolated from single and mixed cultures of Lactobacillus . The same amplification protocol was successfully used directly adding whole bacterial cells to PCR reaction tubes. The suitability of this method for in vivo studies was investigated through feeding L. gasseri 4B2 to mice and analysing colony forming units obtained by plating faecal samples on selective medium. The methodology allows a fast and reliable identification of the target strain without any DNA extraction procedure.