Biomedical subjects

Patricia Garrido

Publications and source records attributed to Patricia Garrido.

3 recordsLinked to original sources

PubMed2005

STEC-EPEC oligonucleotide microarray: a new tool for typing genetic variants of the LEE pathogenicity island of human and animal Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains.

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) are important emerging pathogens that can cause a severe and sometimes fatal illness. Differentiation of eae, tir, espA, espD, and espB gene variants of the locus of enterocyte effacement (LEE) pathogenicity island represents an important tool for typing in routine diagnostics as well as in pathogenesis, epidemiologic, clonal, and immunologic studies. METHODS: Type-specific oligonucleotide microarrays and a PCR scheme were designed and constructed for the detection and typing of genetic variants of the LEE genes. Oligonucleotide probes were tested for their specificity against the corresponding type strain by microarray hybridization using fluorescent DNA, either PCR-amplified (single, multiplex, long-range), chromosomal, or amplified chromosomal DNA. RESULTS: The PCR scheme and the oligonucleotide microarray allowed us to distinguish 16 variants (alpha1, alpha2, beta1, beta2, gamma1, gamma2/theta, delta/kappa, epsilon, zeta, eta, iota, lambda, mu, nu, xi, omicron) of the eae gene, 4 variants (alpha1, beta1, gamma1, gamma2/theta) of the tir gene, 4 variants (alpha1, beta1, beta2, gamma1) of the espA gene, 3 variants (alpha1, beta1, gamma1) of the espB gene, and 3 variants (alpha1, beta1, gamma1) of the espD gene. We found a total of 12 different combinations of tir, espA, espB, and espD genes among the 25 typed strains. CONCLUSIONS: The PCR scheme and the oligonucleotide microarray described are effective tools to rapidly screen multiple virulence genes and their variants in E. coli strains isolated from human and animal infections. The results demonstrate the great genetic diversity among LEE genes of human and animal STEC and EPEC strains.

Animals↗

PubMed2004

Streptococcus equi subsp. ruminatorum subsp. nov., isolated from mastitis in small ruminants.

Six isolates of an unknown Gram-positive, catalase-negative, chain-forming, coccus-shaped organism isolated from ovine and caprine mastitis were characterized by phenotypic and molecular taxonomic methods. On the basis of cellular morphology and the results of biochemical tests, the organism was tentatively identified as a streptococcal species. Comparative 16S rRNA gene sequencing studies confirmed that the organism is a member of the genus Streptococcus, with Streptococcus equi as its closest phylogenetic relative (98.8 % similarity). DNA-DNA pairing studies showed that the unidentified organism displayed more than 70 % relatedness to the type strains of S. equi subsp. equi and subsp. zooepidemicus. Despite the relatively high DNA-DNA reassociation values, biotyping and ribotyping allowed clear differentiation of the unknown bacterium from the two recognized subspecies of S. equi. On the basis of phenotypic and molecular genetic evidence, it is proposed that the unknown Streptococcus isolates from ovine and caprine mastitis be classified as a novel subspecies, Streptococcus equi subsp. ruminatorum subsp. nov. The type strain is CECT 5772(T) (=CCUG 47520(T)=Mt 167(T)).

Animals↗

PubMed2003

Sternal cleft: a surgical opportunity.

PURPOSE: The aim of this study was to evaluate the results from the surgical techniques utilized to repair congenital sternal cleft. METHODS: From January, 1987 to January, 2001, 5,182 patients were seen for chest wall malformations. Eight (0.15%) had sternal cleft. The age at presentation ranged from 15 days to 5 years. Six were girls (75%). The associated malformations were congenital cardiac malformations (2 patients), maxillofacial hemangioma (1 patient). All of them underwent a surgical repair, which could be classified into 3 methods: group 1 had primary closure of the defect (3 patients); group 2 underwent partial resection of the first, second, and third costal cartilages, disruption of the sternoclavicular junction, and closure of the sternal bars with stainless steel wire (3 patients); and group 3 had mobilization and approximation of the sternocleidomastoid muscles with closure achieved with costal homograft and prosthetic mesh (2 patients). The interval for postoperative follow-up was 1 to 8 years. RESULTS: Group 1 patients developed well, although 2 of them had a slight degree of pectus excavatum in the long term not requiring surgical correction. Group 2 Patients developed without problems in all cases. One of the patients from group 3 had unsatisfactory aesthetic and functional results. He underwent reoperation with the second technique, achieving an improved result. CONCLUSIONS: Primary closure of the sternal cleft is the easiest technique. It should be performed in young infants. In the long term it can lead to a mild degree of pectus excavatum. The costal cartilage resection with mobilization of the clavicle achieved excellent results and allowed ready approximation of both sternal halves avoiding the use of costal grafts and prosthetic material.

Age Factors↗