Evaluation of alkaline phosphatase-labelled ipaH probe for diagnosis of Shigella infections.
The presence of many enteropathogens which are not easily detectable by routine stool culture has led to the development of alternative diagnostic methods. One of these techniques, nucleic acid probe hybridization, has been used to identify Shigella spp. and enteroinvasive Escherichia coli (EIEC) in stool specimens through the detection of genetic material encoded by a specific large approximately 200-kbp virulence-related plasmid. In the present study, an alkaline phosphatase-labelled oligonucleotide probe developed to detect the gene for ipaH, a repetitive genetic sequence thought to be present on both the virulence-related plasmid and the chromosomes of all strains of Shigella and EIEC, was tested in a developing-country setting through a prospective clinical trial. In a group of 219 Peruvian adults and children with acute gastroenteritis, the ipaH probe detected 85% of cases of shigellosis and demonstrated a specificity of 95% when compared with simultaneous detection by several stool culture techniques. Additionally, three cases of EIEC infection which could not be diagnosed by culture methods alone were detected with the ipaH probe and were confirmed by plasmid analysis and Sereny testing. These preliminary results suggest that, with further research, the ipaH probe should prove to be a useful and rapid adjunct in the diagnosis of acute gastroenteritis in developing countries.
Department of International Health, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205.
R A
RA
Oberhelman
D J
DJ
Kopecko
M M
MM
Venkatesan
E
E
Salazar-Lindo
E
E
Gotuzzo
A
A
Yi
E
E
Chea-Woo
R
R
Ruiz
C
C
Fernandez-Prada
R
R
León-Barúa
D000469:Alkaline Phosphatase; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004405:Dysentery, Bacillary / Q000175:diagnosis*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012091:Repetitive Sequences, Nucleic Acid; D012760:Shigella / Q000235:genetics* / Q000302:isolation & purification
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D015345:Oligonucleotide Probes; C063500:ipaH protein, Shigella flexneri; D000469:Alkaline Phosphatase