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PubMed2004

Multilocus sequence typing scheme for bacteria of the Bacillus cereus group.

In this study we developed a multilocus sequence typing (MLST) scheme for bacteria of the Bacillus cereus group. This group, which includes the species B. cereus, B. thuringiensis, B. weihenstephanensis, and B. anthracis, is known to be genetically very diverse. It is also very important because it comprises pathogenic organisms as well as bacteria with industrial applications. The MLST system was established by using 77 strains having various origins, including humans, animals, food, and soil. A total of 67 of these strains had been analyzed previously by multilocus enzyme electrophoresis, and they were selected to represent the genetic diversity of this group of bacteria. Primers were designed for conserved regions of housekeeping genes, and 330- to 504-bp internal fragments of seven such genes, adk, ccpA, ftsA, glpT, pyrE, recF, and sucC, were sequenced for all strains. The number of alleles at individual loci ranged from 25 to 40, and a total of 53 allelic profiles or sequence types (STs) were distinguished. Analysis of the sequence data showed that the population structure of the B. cereus group is weakly clonal. In particular, all five B. anthracis isolates analyzed had the same ST. The MLST scheme which we developed has a high level of resolution and should be an excellent tool for studying the population structure and epidemiology of the B. cereus group.

Alleles↗
PubMed1999

Characterization and locus assignment of two alpha-globin variants present in the Maltese population: Hb St. Luke's [alpha95(G2)Pro-->Arg] and Hb Setif [alpha94(G1)Asp-->Tyr].

Two types of alpha-globin variants were found in 0.2% of a large number of newborn from Malta. The two hemoglobins were identified from tryptic maps on a Vydac C18 column and by alpha-globin gene sequencing as Hb St. Luke's (isoelectric point = 7.18+/-0.017) and Hb Setif (isoelectric point = 7.26+/-0.010). Hb St. Luke's [alpha95(G2)Pro-->Arg] was found to result from a C-->G mutation at the second position of codon 95 on an alpha1-globin gene, and Hb Setif [alpha94(G1) Asp-->Tyr] resulted from a G-->T mutation at the first position of codon 94 on an alpha2-globin gene. Quantification of Hb St. Luke's (11.1+/-1.12%) and Hb Setif (14.7+/-2.22%) in peripheral blood hemolysates indicated that, in the absence of either an alpha- or a beta-thalassemia allele, the protein products of the alpha1- and alpha2-globin genes were nearly equal in quantity.

Arginine↗
PubMed1999

Circulation of subtype A and gagA/envB recombinant HIV type 1 strains among injecting drug users in St. Petersburg, Russia, correlates with geographical origin of infections.

Countries of the former Soviet Union are experiencing an emerging HIV-1 epidemic due to a rapid expansion of HIV-1 among injecting drug users (IDUs). To study the molecular epidemiology of HIV-1 among IDUs in St. Petersburg, Russia, virus sequences were obtained from 22 individuals. Phylogenetic analysis of the env and gag regions revealed circulation of two major HIV-1 populations, one belonging to HIV-1 subtype A, and another being a recombinant of subtype A and B viruses (gagA/envB). Both virus populations were highly homogeneous, with a mean pairwise genetic distance of <2%, and similar to viruses obtained earlier from IDUs in other regions of the former Soviet Union. Distribution of the two major HIV-1 genotypes in St. Petersburg correlated with geographical origin of infections. In one individual, a virus type previously unseen among IDUs was found, which demonstrates the possibility that new viruses are entering this risk group.

Adult↗
PubMed2005

Sequence variation in the porB gene from B:P1.4 meningococci causing New Zealand's epidemic.

Since mid-1991, New Zealand has experienced an epidemic of meningococcal disease. The epidemic has been caused by serogroup B meningococci expressing PorA type P1.7-2,4, belonging to the ST-41/ST-44 complex, lineage III. Most B:P1.7-2,4 meningococci express type 4 PorB (87.0%), although case isolates with porB other than type 4 have been identified throughout the duration of the epidemic. To assess the genetic relatedness of case isolates with an alternative porB gene, multilocus restriction typing validated against multilocus sequence typing was used. This determined that B:P1.7-2,4 meningococci with a porB gene that was other than type 4 had the same clonal origin. It was concluded that strains with alternative porB genes had diverged from the original type 4 porB. Variation in porB was also shown to be associated with the uptake of DNA encoding one or two of the PorB variable regions leading to mosaic porB. Point mutation rather than horizontal transfer and recombination was implicated as the mechanism of sequence variation in some strains. This work will serve as a reference point to determine if the administration of a strain-specific vaccine increases the level of porB divergence and variation already observed in New Zealand case isolates. It also complements the study undertaken of PorA stability which showed that variation in P1.7-2,4 PorA was almost exclusively due to deletions in the P1.4 epitope of the epidemic strain.

Amino Acid Sequence↗
PubMed2003

Flavobacterium limicola sp. nov., a psychrophilic, organic-polymer-degrading bacterium isolated from freshwater sediments.

Three novel strains of cold-adapted bacteria, ST-82T, ST-10 and ST-92, were isolated from freshwater sediments. These three isolates were very similar to each other in phenotypic and chemotaxonomic traits, as well as in 16S rDNA sequence. The strains were Gram-negative, elongated filament-like rods that formed bright yellow colonies. They showed neither flexirubin pigments nor gliding motility. The strains were able to hydrolyse casein, gelatin, starch, agar, aesculin, urea, uric acid and tyrosine. They also lysed cells of Escherichia coil and Pseudomonas putida. The temperature range for growth was 0-25 degrees C, with optimum growth occurring at 15-20 degrees C. For all isolates, protease secretion increased as temperature decreased. Sodium chloride inhibited their growth, although the strains tolerated up to 1.5% (w/v) NaCl. Menaquinone-6 was the major respiratory quinone. The major cellular fatty acids were C15 : 0, iso-C15 : 0, anteiso-C15 : 0, C15:1, iso-C15:1, C16 : 1omega7cis, iso-C16 : 1, iso-C17 : 1, iso-C15 : 3-OH and iso-C16 : 0 3-OH. The DNA G + C content was 34.0-34.8 mol%. Phylogenetic analysis based on 16S rDNA sequences suggested that the strains belonged to the genus Flavobacterium and were closely related to Flavobacterium xanthum and Flavobacterium frigidarium, with sequence similarities of 96.9 and 96.3%, respectively. In physiological and biochemical analyses, the isolates were differentiated from all known members of the genus Flavobacterium. The name Flavobacterium limicola is proposed for these novel strains, and the type strain is ST-82T (=JCM 11473T =DSM 15094T).

Base Composition↗
PubMed1989

Comparison of hemodynamic, electrocardiographic, mechanical, and metabolic indicators of intraoperative myocardial ischemia in vascular surgical patients with coronary artery disease.

To compare mechanical, electrocardiographic, and metabolic indices of myocardial ischemia, the cardiokymogram (CKG), the V5 ECG, left anterior descending coronary artery territory lactate extraction, and pulmonary capillary wedge pressure (PCWP) were measured in 53 vascular surgical patients with coronary artery disease. Measurements were performed preoperatively and at four specific intraanesthetic intervals: after tracheal intubation, before surgery, and 10 and 30 min after incision. Measurements and sampling sequence took 5-7 min, and therapy for the probable cause of ischemia was instituted following completion of this sequence. Myocardial ischemia was defined as type II or III CKG, 0.1 mV or greater horizontal or downsloping depression of V5 ECG ST segment, 0.2 mV or greater elevation of V5 ECG ST segment, or myocardial lactate production. Thirty-nine patients (74%) had a total of 89 episodes of myocardial ischemia. Seventy-four episodes (83%) were detected by the CKG, 31 (44%) were evident on the ECG, and 13 (15%) by evidence of lactate production. The concordance among the indices of myocardial ischemia was poor. Patients with an abnormal preoperative ECG experienced a greater number of ischemic episodes (P less than 0.001). Elevation of PCWP or the presence of A-C or V-waves greater than 5 mmHg above the mean did not individually reflect ischemia reliably. Intraoperative myocardial ischemia is common in vascular surgical patients and is most sensitively detected by ventricular wall motion abnormality.

Aged↗
PubMed2005

Inducible clindamycin resistance and molecular epidemiologic trends of pediatric community-acquired methicillin-resistant Staphylococcus aureus in Dallas, Texas.

Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infection occurs commonly in children. Clindamycin resistance may be inducible or constitutive, and the rates of inducible resistance in CA-MRSA that could produce clindamycin treatment failures vary worldwide. The double-disk test was performed in 197 erythromycin-resistant and clindamycin-susceptible CA-MRSA strains from children in Dallas, Texas, from 1999 to 2002 to determine inducible clindamycin resistance. Resistance mechanisms were studied by PCR; epidemiologic trends were studied by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Inducible resistance was demonstrated in 28 (93%+/-6%) of 30 tested isolates in 1999, 21 (64%, +/-11%) of 33 in 2000, 12 (23%+/-7%) of 52 in 2001, and 6 (7%+/-3%) of 82 in 2002. All noninducible strains had the msr(A) gene. Among inducible resistant strains, 31 had erm(B), 24 had erm(C), and 12 had erm(A) genes. Two distinct pulsed types were the most prevalent; one of them was the most common pulsed type in 1999, whereas in 2002 a different pulsed type was prevalent. MLST analyses determined that ST-8 was the most common type, with 76%+/-5% found in 2002. All but one of these clindamycin-susceptible, erythromycin-resistant ST-8 strains showed no induction of clindamycin resistance. We conclude that, among erythromycin-resistant, clindamycin-susceptible CA-MRSA strains isolated from children in Dallas, inducible methylase resistance became less common from 1999 to 2002 (P<0.001). The phenotype of strains was associated with their sequence type. Our results demonstrate a clonal shift in CA-MRSA in Dallas children from 1999 to 2002.

Adolescent↗
PubMed2006

Genetic relatedness between group B streptococci originating from bovine mastitis and a human group B Streptococcus type V cluster displaying an identical pulsed-field gel electrophoresis pattern.

Twenty isolates of group B streptococcus (GBS) were recovered from the milk of cows with bovine mastitis on three farms located in the south and south-east of Brazil between 1987 and 1988. These isolates were characterised by molecular methods and compared with a collection of 103 human GBS isolates from colonised and infected patients in the same region between 1980 and 2003. Some of the bovine isolates shared identical or similar pulsed-field gel electrophoresis (PFGE) patterns with a PFGE clone of human GBS type V. In addition, these bovine and human isolates also possessed the same ribotype. Multilocus sequence typing (MLST) of representative isolates confirmed the genetic relationship between the human and bovine GBS isolates with identical PFGE patterns, which clustered in the same ST-26 clonal complex. These data support the hypothesis that some bovine GBS strains are related closely to human isolates and may infect humans, or vice versa. Further comparative genomic analyses of GBS isolates from bovine and human origins are required to investigate this hypothesis further.

Animals↗
PubMed2026

Genomic and One Health insights into Vibrio parahaemolyticus from environmental, seafood and clinical sources.

Vibrio parahaemolyticus is a leading cause of seafood-borne gastroenteritis worldwide, with climate warming facilitating its spread to high-latitude areas. In this study, we analyzed 212 genomes of environmental and seafood-associated isolates collected from seven cities in Zhejiang Province, China (2019-2024), alongside 228 clinical genomes from public databases. The 212 isolates were assigned to 172 sequence types (STs), with ST490 being the most frequent (5/212, 2.36%). Forty-four serotypes were identified, dominated by OL3:KUT (12.68%). High ST and serotype diversity were observed across different sample types and sources, with median pairwise single nucleotide polymorphisms (SNPs) ranging from 57,431 to 58,378, indicating comparable genetic diversity across groups. All isolates carried tlh and T3SS1 but lacked tdh and T3SS2. Resistance rates against ampicillin and cefazolin were 54.72% (116/212) and 44.34% (94/212), respectively, with multidrug resistance (MDR) detected in nine isolates, predominantly from seafood (7/9). A total of 63 distinct antimicrobial resistance genes (ARGs) spanning seven classes were identified. Isolates from aquaculture farms and wet markets exhibited greater resistance category diversity and higher ARG carriage than those from coastal or riverine sites. In contrast, the 228 clinical isolates harbored only 25 ARGs across two classes, with a significantly lower proportion of isolates carrying multiple ARG classes (0.44% vs. 6.13%, P&#xa0;<&#xa0;0.001). Human isolates formed tighter phylogenetic clusters, although a minority were closely related to environmental/foodborne strains. Overall, our findings demonstrate the genetic diversity and resistance potential of V. parahaemolyticus across environmental, seafood, and clinical sources, highlighting the importance of the One Health approach to comprehensive public health risk assessment.

Vibrio parahaemolyticus↗
PubMed1998

Differentiation-dependent expression of alpha-2,3-sialyltransferase in rabbit corneal epithelium.

PURPOSE: Lectin studies have shown that in the rabbit corneal epithelium, alpha-2,3 sialylation of O-linked glycans differentiates limbal and corneal epithelial cell phenotypes. Because sialic acid can be regulated at the level of the expression of sialyltransferases (STs), the purpose of the present study was to analyze the expression of alpha-2,3STs in this tissue. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to generate ST cDNA from total rabbit corneal epithelium RNA using primers selected from the sequences of three previously cloned STs capable of catalyzing the transfer of sialic acid to O-linked oligosaccharides, human placental Galbeta-1,3GalNAc-Galbeta-1,4GluNAcalpha-2,3ST (STZ), and mouse brain Galbeta-1,3GalNAcalpha-2,3ST types I and II (ST3Gal I and ST3Gal II). Tissue distribution of mRNA was assayed by fluorescent in situ hybridization. A synthetic peptide whose sequence was deduced from a cloned cDNA fragment was synthesized and used to prepare an anti-ST goat antiserum. The molecular weights of immunodetectable polypeptides and their distribution in cryostat sections of the limbocorneal area were investigated by western blot analysis and indirect immunofluorescence, respectively. RESULTS: RT-PCR yielded cDNA of expected basepair length for STZ and ST3(Gal II. The rabbit STZ cDNA was 86% identical with its human equivalent. Its mRNA was confined to the cornea, mainly in basal epithelial cells, and was not expressed in the limbus. Western blot analysis identified a band at 37 kDa whose binding was abolished by preincubation of the antiserum with the immunization peptide. Immunohistologic analysis revealed the presence of immunoreactive epitopes in all basal cells of the cornea but not in the limbus. CONCLUSIONS: STZ mRNA and the enzyme itself are expressed in the basal layer of the corneal epithelium but are absent in the limbus. This enzyme's de novo expression seems thus responsible for the differential expression of alpha-2,3 sialylation along the limbocorneal differentiation axes. At least one more alpha-2,3ST is also present in the epithelium.

Animals↗
PubMed1993

Rapid identification of flavivirus using the polymerase chain reaction.

A rapid and accurate detection and identification system was developed for flaviviruses that makes use of reverse transcription-polymerase chain reaction (RT-PCR). A primer pair (YF-1 and YF-3), which corresponds to the highly conserved sequence at 3' noncoding region among flaviviruses, was useful for identification of mosquito-borne flaviviruses. Nine sets of species specific primer pairs were also selected to identify and distinguish species, i.e., yellow fever, West Nile, Murray Valley encephalitis, Japanese encephalitis, St. Louis encephalitis and dengue type 1 to 4 viruses. This method required only two hours for completion using infected culture fluid, thus facilitating rapid identification of mosquito-borne flavivirus species.

Animals↗
PubMed2006

Enhancer- and silencer-like sequences that mediate insulin-like growth factor-binding protein-2 gene expression in uterine cells of pregnancy.

The regulation of insulin-like growth factor-binding protein-2 (IGFBP-2) gene transcription in specific cell and developmental contexts is not well understood. Here, we identified DNA regions that mediate IGFBP-2 gene transcription in two of the three major cell types of the uterine endometrium of the early pregnant pig. Two clusters of transcriptional start sites at nucleotides -109/-105 and -96/-87 (+1, translational initiation site) in the porcine IGFBP-2 gene were localized in uterine endometrium and in primary cultures of endometrial glandular epithelial (GE) and stromal (ST) fibroblastic cells. Upstream regions of this gene (spanning -1,397/+73) were fused to a luciferase reporter gene, and the constructs were transiently transfected into endometrial GE and ST cells representative of pregnancy days 12 and 18 (day 115 = parturition). A short (110 bp) upstream region (-874/-765) stimulated the IGFBP-2 and heterologous SV40 promoters in the two cell types at both pregnancy days. Two noncontiguous copies of the novel sequence motif TCAGGG within the 110-bp fragment were implicated in transcriptional activity, since block mutation of these sequences led to a repression of SV40 basal promoter activity in endometrial cells. Southwestern blotting identified an endometrial nuclear protein of 34-kDa molecular weight that bound an oligonucleotide containing this motif, and EMSA suggested robust expression of this protein in early pregnancy endometrium and in ovary but at much reduced levels in endometrium at later pregnancy. A pair of E-box elements (CANNTG) within the 110 bp region was stimulatory to IGFBP-2 promoter activity; block mutation of these converted the 110-bp region into a potent transcriptional silencer in all but day 18 ST cells. Results identify novel DNA motifs that regulate the IGFBP-2 gene promoter in uterine endometrium in pregnancy-associated fashion.

Animals↗
PubMed2005

Gene sequence heterogeneity of Corallococcus coralloides strains isolated from geographically diverse locations.

Thirty-three strains classified as Corallococcus coralloides isolated from mostly soil samples in 14 countries of four continents, were subjected to phylogenetic analyses. Based on 16S rDNA analyses the strains form a highly related cluster, sharing above 98.7% sequence similarity. Four groups were recognized within this cluster, only one of which, containing two strains from St. Lucia, Lower Antilles, was exclusively defined by strains from the same sample. The other groups contained members from different countries, even continents. The largest group embraced the type strains of C. coralloides DSM 2259(T) and Corallococcus exiguus 14696(T) which were almost indistinguishable in their 16S rRNA gene sequence. Corallococcus macrosporus DSM 14697(T) grouped outside the C. coralloides cluster, showing a higher relationship to a member of Myxococcus. The topology of the tree generated on the basis of the partial gyrase B (gyrB) gene sequence supports the rRNA gene tree, though some differences in the order of branching were observed. As judged by the binary similarity values the higher resolution power of gyrB sequences was confirmed. From a taxonomic standpoint, the size of myxospores is not a valuable taxonomic criterion, as small- and medium-sized myxospores are members of the same group. If the species status of C. coralloides and C. exiguus is verified by other methods (e.g. DNA-DNA hybridisation, RiboTyping), the genus Corallococcus may embrace a broad range of yet-to-be described novel species. The presence of strains within the same sample displaying higher relatedness to strains from other locations points towards an intensive dispersal of myxospores across continents.

Asia↗
PubMed2003

Mechanism of ST elevation and ventricular arrhythmias in an experimental Brugada syndrome model.

BACKGROUND: Although phase 2 reentry is said to be responsible for initiation of ventricular tachycardia (VT) in Brugada syndrome, information about the activation sequence during VT is limited. METHODS AND RESULTS: We developed an experimental Brugada syndrome model using a canine isolated right ventricular preparation cross-circulated with arterial blood of a supporter dog and examined the VT mechanism. Two plaque electrodes (35x30 mm) containing 96 bipolar electrodes were attached to the endocardium and epicardium. Saddleback and coved types of ST elevation in transmural ECG were induced by pilsicainide, a pure sodium channel blocker, and pinacidil, a KATP channel opener. Eighteen polymorphic VT episodes were recorded in 9 of the 12 preparations associated with ST elevation. Fourteen episodes spontaneously developed in 5 preparations after an extrasystole during basic drive pacing. Analysis of local recovery times revealed increased dispersion especially in epicardium, and the extrasystole originated from a site with a short recovery time, suggesting that phase 2 reentry was its mechanism. The other 4 VTs in 4 preparations were induced by premature stimulation. Analysis of the activation sequences during VT revealed reentry between epicardium and endocardium or reentry around an arc of a functional block confined to epicardium or endocardium with bystander activation of the other. CONCLUSIONS: Electrical heterogeneity in the recovery phase was induced in this experimental Brugada syndrome model, which can be a substrate for the development of phase 2 reentry and the subsequent reentry around an arc of the functional block, resulting in sustained VT.

Animals↗
PubMed2005

Comparison of genetic backgrounds of methicillin-resistant and -susceptible Staphylococcus aureus isolates from Portuguese hospitals and the community.

In order to understand the origins of the dominant methicillin-resistant Staphylococcus aureus (MRSA) clones in Portuguese hospitals, we compared the genetic backgrounds of nosocomial MRSA with methicillin-susceptible S. aureus (MSSA) isolates from the same hospitals (n=155) and from the community (n=157) where they were located. Pulsed-field gel electrophoresis, spa typing, multilocus sequence typing, and agr type analysis revealed that the genetic backgrounds correspondent to the dominant MRSA clones in Portuguese hospitals during the last 15 years (Iberian ST 247, Brazilian ST 239, and EMRSA-15 ST 22) were scarcely or not found among the present MSSA collection. The four major MSSA clones encountered (A-ST 30, B-ST 34, C-ST 5, and H-ST 45) correspond, or are very similar, to the background of other international MRSA pandemic clones, i.e., EMRSA-16, New York/Japan, Pediatric, and Berlin clones. However, with the exception of the Pediatric clone, none of these MRSA clones has been detected in Portugal. Our findings suggest the three major MRSA clones identified in Portuguese hospitals have not originated from the introduction of SCCmec into dominant MSSA backgrounds present in the Portuguese nosocomial or community environment but were probably imported from abroad. In contrast, the MRSA Pediatric clone might have originated in our country by the acquisition of SCCmec type IV into MSSA clone C. Furthermore, we provide evidence that the introduction of SCCmec into sensitive clones is most likely a relatively infrequent event that seems to depend not exclusively on the presence of a successful MSSA lineage.

Anti-Bacterial Agents↗
PubMed2003

Three new antibiotic producing species of the genus Amycolatopsis, Amycolatopsis balhimycina sp. nov., A. tolypomycina sp. nov., A. vancoresmycina sp. nov., and description of Amycolatopsis keratiniphila subsp. keratiniphila subsp. nov. and A. keratiniphila subsp. nogabecina subsp. nov.

Screening of Indian soils for strains producing antibiotics active against methicillin resistant staphylococci resulted in the isolation of the actinomycetes FH 1894T, ST 101170T, and FH 1893T, producing novel glycopeptide antibiotics and a polyenic compound, respectively. Studies of the cellular fatty acids and the absence of mycolic acids as well as the 16S rRNA gene sequence analysis of the producers indicated their membership to the genus Amycolatopsis. Comparison of genomic and metabolic properties of these strains with known species of this genus indicated that they represent new species for which the names Amycolatopsis balhimycina (type strain FH 1894T, DSM 44591T) and Amycolatopsis vancoresmycina (type strain ST 101170T, DSM 44592T) are proposed. The producer of the antibiotic Tolypomycin, strain IFO 14664T, representing the invalid species "Amycolatopsis tolypophorus", was also included in this investigation. As this strain was found to represent a valid species of the genus Amycolatopsis the name Amycolatopsis tolypomycina is proposed for this species, with the type strain IFO 14664T (DSM 44544T). The producer of the antibiotic Nogabecin, strain FH 1893T, was found to be closely related to the type strain of A. keratiniphila DSM 44409T which justifies the description of two subspecies of this species, A. keratiniphila subsp. keratiniphila DSM 44409T and A. keratiniphila subsp. nogabecina DSM 44586T.

Actinomycetales↗
PubMed2002

Denaturing gradient gel electrophoretic multilocus sequence typing of Staphylococcus aureus isolates.

To obviate the need for multilocus sequencing, a method using denaturing gradient gel electrophoresis (DGGE) was developed for the multilocus sequence typing (MLST) of Staphylococcus aureus isolates. Sequence types (STs) were obtained on the basis of sequences of polymerase chain reaction (PCR) products from seven housekeeping genes and compared to the reference MLST database. The melt curves, sequences and DGGE profiles were compared for 100 STs (i) to determine PCR conditions with 40-mer GC-clamps attached to the forward and reverse primers; (ii) to choose single restriction enzyme sites for digestion of PCR products into two fragments each with a GC-clamp attached and (iii) to optimize DGGE conditions. When the DGGE types (DT) were analyzed, the majority of DTs (76/100) were accurately classified into one ST (95% of nucleotide changes were detected), 10 DTs were classified into one of two STs corresponding to a single nucleotide ambiguity and 14 DTs were classified into 3 or 4 STs corresponding to 4 or 5 nucleotide ambiguities. A combination of STs and DTs were used to obtain septuplet sets of STs (7-ST) for 25 S. aureus isolates. When compared to the reference MLST database, one methicillin-resistant S. aureus (MRSA) isolate had the same genotype as the first MRSA clone. The DGGE-MLST method can be used as a rapid, accurate and 20-fold less expensive method than DNA sequencing for the detection of all sequence types. This combined laboratory and in silico approach could have wide applicability not only to MLST methods for other bacteria but to the screening of multilocus nucleotide differences deposited in other mutation databases.

Alleles↗
PubMed2005

Characterization of major clones of antibiotic-resistant Streptococcus pneumoniae in New Zealand by multilocus sequence typing.

OBJECTIVES: To determine the relationship between New Zealand isolates of antibiotic-resistant Streptococcus pneumoniae and internationally widespread resistant clones. METHODS: Fifty-nine isolates representing both invasive and non-invasive pneumococci from multiple locations, serotypes and years were analysed by multilocus sequence typing. RESULTS: Major international clones, including Spain(23F)-1, France(9V)-3 and Taiwan(19F)-14, were found to be present in New Zealand. A one-allele variant of the Taiwan(19F)-14 clone (aroE 15-->4, ST 271) was particularly prominent. CONCLUSIONS: Antibiotic-resistant pneumococci have not evolved de novo in New Zealand, but were introduced to the country during the early 1990s.

Bacterial Typing Techniques↗