scholarly journals Ongoing outbreak of invasive listeriosis, Germany, 2012 to 2015

2015 ◽  
Vol 20 (50) ◽  
Author(s):  
Werner Ruppitsch ◽  
Rita Prager ◽  
Sven Halbedel ◽  
Patrick Hyden ◽  
Ariane Pietzka ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Gel Electrophoresis ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Source Case ◽  
Serotype 1 ◽  
Outbreak Investigations ◽  
Cluster Type ◽  
Almost All ◽  
Generation Sequencing

Listeriosis patient isolates in Germany have shown a new identical pulsed-field gel electrophoresis (PFGE) pattern since 2012 (n = 66). Almost all isolates (Listeria monocytogenes serotype 1/2a) belonged to cases living in southern Germany, indicating an outbreak with a so far unknown source. Case numbers in 2015 are high (n = 28). No outbreak cases outside Germany have been reported. Next generation sequencing revealed the unique cluster type CT1248 and confirmed the outbreak. Investigations into the source are ongoing.

scholarly journals Pulsed-Field Gel Electrophoresis Analysis of Nasopharyngeal Flora in Children Attending a Day Care Center

2000 ◽  
Vol 38 (2) ◽  
pp. 625-629 ◽  
Author(s):  
Hisakazu Yano ◽  
Mitsuko Suetake ◽  
Akio Kuga ◽  
Kazuhiko Irinoda ◽  
Ryoichi Okamoto ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Moraxella Catarrhalis ◽  
Day Care ◽  
Care Center ◽  
Acute Otitis Media ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Day Care Center ◽  
Pulsed Field ◽  
Almost All

To investigate how bacterial pathogens spread from child to child in a day care center, we monitored six children, two boys and four girls, born between August 1995 and November 1997, attending a day care center and analyzed nasopharyngeal samples from them using pulsed-field gel electrophoresis (PFGE). We obtained nasopharyngeal cultures from all of the affected children and almost all of the unaffected children between September 1998 and March 1999 after some children presented simultaneously with purulent rhinorrhea. Moreover, when a child was found to have acute otitis media, nasopharyngeal secretions from the child were independently cultured during treatment. During this period, 28 isolates of Moraxella catarrhalis, 13 ofStreptococcus pneumoniae, and 4 of Haemophilus influenzae were recovered. PFGE gave 8 patterns for M. catarrhalis, 10 for S. pneumoniae, and 1 for H. influenzae. PFGE patterns demonstrated spread of M. catarrhalis between children. However, each occurrence of clusters of infection with M. catarrhalis lasted 2 to 6 weeks, with a change in PFGE pattern between occurrences of clusters. The M. catarrhalis strain infecting each child also changed. Similarly, the S. pneumoniae strain in each child also changed. In contrast, infection with H. influenzaepersisted for about 3 months in an affected child.

Rapid Inactivation of Non-Endospore-Forming Bacterial Pathogens by Heat Stabilization is Compatible with Downstream Next-Generation Sequencing

2019 ◽  
Vol 24 (3) ◽  
pp. 129-133
Author(s):  
Max R. Schroeder ◽  
Vladimir Loparev
Keyword(s):  
Next Generation Sequencing ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
High Density ◽  
Next Generation ◽  
Gram Positive ◽  
Gram Negative ◽  
Pulsed Field ◽  
Stabilization Treatment ◽  
Generation Sequencing

Introduction: Heat stabilization treatment preserves the in vivo state of biological samples by rapidly inactivating enzymes that cause degradation of proteins and nucleic acids. Historically, proteomics studies used this technique as an alternative to chemical fixation. More recently, microbiologists discovered that heat stabilization treatment rapidly inactivates pathogens present in tissue samples and preserves deoxyribonucleic acid (DNA) in the tissue. However, these recent studies did not investigate the inactivation of high-density bacterial suspensions and the quality of bacterial DNA. Methods and Results: High-density suspensions of Escherichia coli (>109 cfu/mL) were completely inactivated by heat stabilization treatment using the Denator Stabilizor T1 instrument at 72°C and 95°C for 45 seconds. Using the heat stabilization instrument, a panel of 30 species, 20 Gram-negative and 10 non-endospore-forming Gram-positive species, were fully inactivated by treatment (95°C for 45 seconds). DNA was isolated from bacterial suspensions of Gram-negative bacteria, including E. albertii, E. coli, Shigella dysenteriae, and S. flexneri, following inactivation via heat stabilization treatment and without treatment. DNA isolated following heat stabilization treatment was fully compatible with all downstream molecular applications tested, including next-generation sequencing, pulsed-field gel electrophoresis, multiplex polymerase chain reaction (PCR), and real-time PCR. Conclusions and Discussion: Heat stabilization treatment of Gram-negative and non-endospore-forming Gram-positive pathogens completely inactivates high-density bacterial suspensions. This treatment is compatible with downstream DNA molecular assays, including next-generation sequencing, pulsed-field gel electrophoresis, and PCR. Inactivation by heat stabilization is a rapid process that may increase safety by decreasing risks for laboratory-associated infections and risks associated with transportation of infectious materials.

Surveillance forListeria monocytogenesand listeriosis, 1995–2004

2009 ◽  
Vol 138 (4) ◽  
pp. 559-572 ◽  
Author(s):  
C. G. CLARK ◽  
J. FARBER ◽  
F. PAGOTTO ◽  
N. CIAMPA ◽  
K. DORÉ ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Patient Characteristics ◽  
Pulsed Field Gel Electrophoresis ◽  
Outbreak Detection ◽  
Pfge Analysis ◽  
Serotype 1 ◽  
Outbreak Investigations ◽  
And Control

SUMMARYCanadian cases and outbreaks of illness caused byListeria monocytogenesbetween 1995 and 2004 were assessed. Isolates (722 total) were characterized by serotyping, and pulsed-field gel electrophoresis (PFGE) was performed to provide a means of detecting case clusters. Rates of listeriosis remained fairly consistent during the period of study, and patient characteristics were similar to those seen in studies of other populations. Most isolates were obtained from blood and cerebrospinal fluid, although during some outbreak investigations isolates were also obtained from stools. Serotype 1/2a predominated in isolates from patients in Canada, followed by serotypes 4b and 1/2b. Outbreaks caused byL. monocytogenesthat occurred during the period of study were caused by isolates with serotypes 1/2a and 4b. A retrospective analysis of PFGE data uncovered several clusters that might have represented undetected outbreaks, suggesting that comprehensive prospective PFGE analysis coupled with prompt epidemiological investigations might lead to improved outbreak detection and control.

scholarly journals Application of pulsed-field gel electrophoresis to the epidemiological characterization ofStaphylococcus intermediusimplicated in a food-related outbreak

1994 ◽  
Vol 113 (1) ◽  
pp. 75-81 ◽  
Author(s):  
F. M. Khambaty ◽  
R. W. Bennett ◽  
D. B. Shah
Keyword(s):  
Gel Electrophoresis ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Western United States ◽  
Single Strain ◽  
Pulsed Field ◽  
Definitive Evidence ◽  
Staphylococcus Intermedius ◽  
Epidemiological Characterization

SUMMARYAn outbreak of food intoxication involving over 265 cases in western United States occurred in October 1991.Staphylococcus intermediuswas implicated as the aetiologic agent. Representative outbreak isolates (five clinical and ten from foods) produced type A enterotoxin. DNA fragments generated by four restriction endonucleases and analysed by pulsed-field gel electrophoresis (PFGE) provided definitive evidence that all isolates from nine different counties in California and Nevada were derived from a single strain. The PFGE pattern of these outbreak isolates was distinct from those of a heterogeneous collection of sevenS. intermediusstrains of veterinary origin and five unrelatedS. aureuslaboratory strains. The data show a significant PFGE pattern heterogeneity not only among members of differentStaphylococcusspecies but also within members of the same species and even the same enterotoxin type. The results indicate that PFGE is a valuable strain-specific discriminator for the epidemiological characterization ofS. intermedius. To our knowledge, this represents the first documented foodborne outbreak caused byS. intermedius. These findings suggest that the presence ofS. intermediusand other species such asS. hyicusin food should be reason for concern.

scholarly journals Detection of Coccidioides posadasii in a patient with meningitis using metagenomic next-generation sequencing: a case report

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yuqiao Mao ◽  
Xia Li ◽  
Haibo Lou ◽  
Xiaoyu Shang ◽  
Yanjun Mai ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Los Angeles ◽  
Systemic Infection ◽  
High Sensitivity ◽  
Pathological Examination ◽  
Next Generation ◽  
Coccidioides Posadasii ◽  
Dimorphic Fungi ◽  
Almost All ◽  
Generation Sequencing

Abstract Background Coccidioidomycosis is a systemic infection caused by dimorphic fungi Coccidioides spp. endemic to Southwestern United States and Central and South America. A history of residence and travel in these areas is essential for the diagnostic of coccidioidomycosis, which has highly variable symptoms ranging from asymptomatic to severe, disseminated infection, and even death. Immunocompromised patients of coccidioidomycosis experience a high risk of dissemination, chronic infection, and mortality. Meningitis is one of the most deleterious coccidioidomycosis and can cause various life-threatening complications. Case presentation Here we report a case of Coccidioides posadasii meningitis in a 49-year-old female who returned to China after one and a half years residence in Los Angeles, USA. The repeated routine cultures using CSF for bacteria or fungi were all negative. To hunt for an infectious etiology, the state-of-the-art technology metagenomic next-generation sequencing (mNGS) was then utilized, suggesting Coccidioides posadasii. Organizational pathological examination and polymerase-chain-reaction (PCR) results subsequently confirmed the mNGS detection. Conclusion To our knowledge, cases for coccidioidal meningitis have been rarely reported in China. While global travelling may spread this disease across continents and make the diagnosis more difficult. mNGS can detect almost all known pathogens with high sensitivity and specificity, especially for uncommon pathogen, such as Coccidioides posadasii in China.

scholarly journals Next generation sequencing of human enterovirus strains from an outbreak of enterovirus A71 shows applicability to outbreak investigations

2020 ◽  
Vol 122 ◽  
pp. 104216
Author(s):  
Sacha Stelzer-Braid ◽  
Matthew Wynn ◽  
Richard Chatoor ◽  
Matthew Scotch ◽  
Vidiya Ramachandran ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Human Enterovirus ◽  
Next Generation ◽  
Enterovirus A71 ◽  
Outbreak Investigations ◽  
Generation Sequencing

scholarly journals Molecular Epidemiology of Penicillin-ResistantStreptococcus pneumoniae Isolates Recovered in Italy from 1993 to 1996

1998 ◽  
Vol 36 (10) ◽  
pp. 2944-2949 ◽  
Author(s):  
Anna Marchese ◽  
Mario Ramirez ◽  
Gian Carlo Schito ◽  
Alexander Tomasz
Keyword(s):  
Gel Electrophoresis ◽  
Fragment Length Polymorphism ◽  
Antibiotic Resistance Genes ◽  
Northern Italy ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Type ◽  
Penicillin Binding Protein ◽  
Additional Resistance ◽  
Restriction Fragment Length

Thirty-nine penicillin-resistant Streptococcus pneumoniae isolates recovered among the approximately 700 pneumococcal strains collected from 1993 to 1996 in central and northern Italy were analyzed for several characteristics, including serotype, antibiotic susceptibility profile, chromosomal relatedness (by using pulsed-field gel electrophoresis [PFGE]), restriction fragment length polymorphism (RFLP) of the penicillin-binding protein (PBP) genes 1A, 2X, and 2B, and the presence of a variety of antibiotic resistance genes (determined by hybridization with appropriate DNA probes). The MICs of penicillin for most of the isolates (30 of 39) were high, in the range of 1 μg/ml or higher, and these 30 isolates carried additional resistance traits to two or more drugs (erythromycin, chloramphenicol, co-trimoxazole, and tetracycline) and expressed serotypes 9, 19, and 23 and three distinct PFGE patterns. More than half (22 of 30) of the isolates for which MICs were high were identified as representatives of two widespread international epidemic clones of S. pneumoniae. The first one of these clones (seven isolates) expressed serotype 23F and possessed all properties characteristic of the widespread Spanish/USA international clone. Seven additional strains with serotype 19 also had the same PFGE pattern, PBP gene, and RFLP polymorphisms, and other properties typical of the serotype 23 Spanish/USA clone, suggesting that these strains were the products of a capsular transformation event (from serotype 23F to serotype 19) in which the Spanish/USA clone was the recipient. The second international clone was represented by eight serotype 9 isolates which were resistant to penicillin and trimethoprim-sulfamethoxazole and had the molecular properties of the French/Spanish epidemic clone. The remaining eight isolates for which penicillin MICs were high appeared to represent a hitherto-undescribed “Italian” clone; they had a novel PFGE type, unique RFLPs for the PBP genes, and resistance to tetracycline, trimethoprim-sulfamethoxazole, and erythromycin, and the penicillin MICs for these isolates were 2 to 4 μg/ml.

Monitoring of Chicken Houses and an Attached Egg-Processing Facility in a Laying Farm for Salmonella Contamination between 1994 and 1998

2001 ◽  
Vol 64 (12) ◽  
pp. 1912-1916 ◽  
Author(s):  
TOSHIYUKI MURASE ◽  
KAZUKO SENJYU ◽  
TAKESHI MAEDA ◽  
MASAYUKI TANAKA ◽  
HIROSHI SAKAE ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Salmonella Enterica ◽  
Environmental Samples ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Drain Water ◽  
Pulsed Field ◽  
Processing Facility ◽  
Single Clone

Two chicken houses and an attached egg-processing facility in a laying farm were sampled between 1994 and 1998 to investigate Salmonella contamination. Each of the houses was environmentally controlled and fitted with egg belts that transported eggs from the houses to the egg-processing facility. Four hundred twenty-eight Salmonella isolates were obtained from 904 environmental samples collected from the houses. Two hundred fifty-two of the 428 (58.9%) isolates yielded five serotypes as follows: Salmonella enterica subsp. enterica serovar Livingstone, Salmonella serovar Cerro, Salmonella serovar Montevideo, Salmonella serovar Mbandaka, and Salmonella serovar Corvallis. The remaining (41.1%, 176 of 428) isolates included four other serotypes and isolates that were untypeable. Salmonella isolates obtained from the drain water collected after the washing of the eggs in the egg-processing facility yielded the same serotypes as those found in the chicken houses. Strains having an identical pulsed-field gel electrophoresis (PFGE) pattern were continually recovered from a house for more than 1 year. Several strains of Salmonella Cerro, Salmonella Mbandaka, and Salmonella Montevideo obtained from both the houses and from the egg-processing facility were indistinguishable by PFGE, respectively. These results suggest that Salmonella organisms originating from a single clone colonized the chicken houses and that the egg belts are likely to be one of the means by which Salmonella organisms are spread from one house to the others.

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