A new multiplex PCR for differential identification of Shigella flexneri and Shigella sonnei and detection of Shigella virulence determinants.

Most of the multiplex PCR (mPCR) used to identify Shigella do not discriminate between Shigella species or serotypes. We designed a mPCR to differentiate between S. flexneri and S. sonnei strains based on the detection of markers associated with the she pathogenicity island described in Shigella. In addition, specific primers were included to detect the Shigella virulence determinants ShET-1 and ShET-2 enterotoxin genes. The analysis of 304 Shigella strains from Chile and 79 Shigella strains from other geographic locations indicated that the mPCR described here detected all Shigella species and specifically differentiated S. flexneri and S. sonnei. The technique was sensitive, reproducible, specific and simple to perform, providing a new tool with the potential to be employed for epidemiological and diagnostic purposes.

Detection of enterohemorrhagic Escherichia coli in meat foods using DNA probes, enzyme-linked immunosorbent assay and polymerase chain reaction.

Enterohemorrhagic Escherichia coli (EHEC) is an important cause of diarrhoea with blood and haemolytic uremic syndrome (HUS) in children and elderly people. Infections with EHEC are a world-wide public health problem, related to consumption of contaminated ground beef. The aim of this study was to establish whether different meat foods sold in Santiago, Chile pose an infection risk by EHEC and to evaluate three different diagnostic techniques in foods, to determine which is most applicable for use in Chile. A parallel analysis was performed on 64 samples of meat foods (23 refrigerated ground meat, 23 refrigerated long pork sausages and 18 frozen hamburgers) sold in Santiago, Chile using DNA probes, enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Twenty-four samples (24 of 64 = 37.5%) were positive by DNA probes, ELISA or PCR. The positive and negative predictive values, sensitivity and specificity of ELISA were 26.7, 81.6, 30.8 and 78.4%, respectively. The positive and negative predictive values, sensitivity and specificity of PCR were 91.7, 96.2, 84.6 and 98%, respectively. The EHEC serogroups most frequently isolated were O158, O157, O119, O125 and O114. These results show that, although molecular techniques such as enzyme immunoassays are useful for EHEC detection in meat foods, PCR has advantages in terms of sensitivity, specificity, cost and ease of implementation in Chile.

[Detection of pyrogenic exotoxin SpeA, SpeB and SpeC genes in Chilean streptococci isolates and their association with clinical manifestations]. / Detección de los genes de las exotoxinas pirogénicas SpeA, SpeB y SpeC en cepas chilenas de Streptococcus pyogenes y su asociación con la clínica.

BACKGROUND: The virulence of Streptococcus pyogenes is determined by a variety of structural molecules, toxins and complex enzymes. Pyrogenic exotoxins cause fever, erythematous reactions, cytotoxic and immunological effects. AIM: To assess the frequency of speA, SpeB and SpeC genes in Chilean Streptococcus pyogenes strains and their association with the invasiveness of infections. MATERIAL AND METHODS: The genes for pyrogenic exotoxins SpeA, SpeB and SpeC were determined by polymerase chain reactions in 114 strains of group A Streptococcus pyogenes isolated from Chilean patients with invasive or non invasive infections. RESULTS: The gene for SpeA was present in 30.7% of isolates, the gene for SpeB was present in 69.3% and the gen for SpeC in 44.7% of isolates. The gene for SpeA was present in 20 of 33 invasive infections and in 15 of 81 non invasive infections (p < 0.0001). On the contrary, the gene for SpeC was present in 11 of 33 invasive infections and in 41 of 81 non invasive infections (p < 0.05). The frequency of speB was similar in invasive and non invasive infections. CONCLUSIONS: There is a clear relationship between the presence of SpeA genes and the severity of infections caused by Streptococcus pyogenes.

[Characterization of a multiresistant strain of Vibrio cholerae O1, isolated from a case of cholera in Chile]. / Caracterización de una cepa de Vibrio Cholerae O1 multiresistente, aislada de un caso de cólera en Chile.

This report characterizes a multiresistant Vibrio Cholerae O1 strain, isolated from a patient with cholera, and investigates the mechanism of resistance. The analyzed strain was resistant to tetracycline, chloramphenicol and trimethoprim-sulfamethoxazole. The resistance was mediated by a 101 megadalton plasmid that was transferred to the resultant of a conjugation assay between the multiresistant V. Cholerae strain and E. coli C-600 used as receptor strain, that acquired the triple resistance of the parental strain. The resistant V. cholerae strain had a Ogawa serotype, El Tor biotype and toxigenic capacity, demonstrated by ELISA and latex agglutination techniques. The biochemical features of the strain were identical to those of susceptible strains, except for the resistance to 10 and 150 ug o 129 vibriostatic factor. The emergence of plasmid mediated resistance to drugs of choice in the treatment of cholera must alert Chilean and Latin American health authorities, considering the cholera will continue affecting the region.

[Characterization of Neisseria meningitidis isolated fron systemic infections. Chile, 1992-1993]. / Caracterización de Neisseria meningitidis aislada de infecciones sistémicas. Chile, 1992-1993.

BACKGROUND: in Chile, all systemic infections caused by Neisseria meningitidis must be reported and the bacterial strain must be sent to a Reference Laboratory at the Instituto de Salud Pública de Chile (ISP). AIM: to report the characterization of strains of N. meningitidis isolated during systemic infections in Chile during the years 1992 and 1993. METHODS: the serogroup, serotype, subtype and antimicrobial susceptibility of every strain of N. meningitidis received at the ISP during 1992 and 1993 was studied. RESULTS: six hundred twenty eight strains of N. meningitidis were confirmed during 1992 and 1993. B serogroup was responsible of 91.1% and 94.7% of confirmed cases during 1992 and 1993 respectively. Serotypes and subtypes most frequently associated to B serogroup were B: 15: P1.3 (63.2%) in 1992 and 51.8% in 1993) and B:NT:P1.3 (11.7% in 1992 and 21.3% in 1993). In 1992, all strains were susceptible to penicillin, chloramphenicol, ceftriaxone and rifampicin. During 1993, 7 (2%) strains were found, for the first time in Chile, moderately susceptible to penicillin and rifampicin MIC90 increased fourfold in respect of 1992, although all strains continued to be susceptible to this antimicrobial. CONCLUSIONS: the increasing frequency of NT (non typified strains) isolation will demand the use of molecular biology techniques for their identification. The appearance of penicillin resistant strains in our country is worrisome.