scholarly journals Development of a Colloidal Gold-Based Immunochromatographic Strip for Rapid Detection of Klebsiella pneumoniae Serotypes K1 and K2

2016 ◽  
Vol 54 (12) ◽  
pp. 3018-3021 ◽  
Author(s):  
L. Kristopher Siu ◽  
Yu-Kuo Tsai ◽  
Jung-Chung Lin ◽  
Te-Li Chen ◽  
Chang-Phone Fung ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Colloidal Gold ◽  
Capsular Polysaccharide ◽  
Polyclonal Antibodies ◽  
Bacterial Species ◽  
False Negative ◽  
Simultaneous Detection ◽  
Test Methods ◽  
Immunochromatographic Strip ◽  
Content Type

In this study, a novel colloidal gold-based immunochromatographic strip (ICS) containing anti-Klebsiella pneumoniaecapsular polysaccharide polyclonal antibodies was developed to specifically detectK. pneumoniaeserotypes K1 and K2. Capsular polysaccharide K1 and K2 antigens were first used to produce polyclonal anti-K1 and anti-K2 antibodies. Reference strains with different serotypes, nontypeableK. pneumoniaestrains, and other bacterial species were then used to assess the sensitivity and specificity of these test strips. The detection limit was found to be 105CFU, and the ICSs were stable for 6 months when stored at room temperature. No false-positive or false-negative results were observed, and equivalent results were obtained compared to those of more conventional test methods, such as PCR or serum agglutination. In conclusion, the ICS developed here requires no technical expertise and allows for the specific, rapid, and simultaneous detection ofK. pneumoniaeserotypes K1 and K2.

scholarly journals Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of Klebsiella pneumoniae Causing Neonatal Sepsis

Pharmaceutics ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1141
Author(s):  
Noha M. Elhosseiny ◽  
Tamer M. Samir ◽  
Aliaa A. Ali ◽  
Amani A. El-Kholy ◽  
Ahmed S. Attia
Keyword(s):  
Gold Nanoparticles ◽  
Klebsiella Pneumoniae ◽  
Causative Agent ◽  
Neonatal Sepsis ◽  
Bacterial Species ◽  
Resistant Bacteria ◽  
Immunochromatographic Strip ◽  
Amino Acid Peptide ◽  
Negative Results ◽  
Drug Resistant Bacteria

Neonatal sepsis is a leading cause of death among newborns and infants, especially in the developing world. The problem is compounded by the delays in pinpointing the causative agent of the infection. This is reflected in increasing mortality associated with these cases and the spread of multi-drug-resistant bacteria. In this work, we deployed bioinformatics and proteomics analyses to determine a promising target that could be used for the identification of a major neonatal sepsis causative agent, Klebsiella pneumoniae. A 19 amino acid peptide from a hypothetical outer membrane was found to be very specific to the species, well conserved among its strains, surface exposed, and expressed in conditions simulating infection. Antibodies against the selected peptide were conjugated to gold nanoparticles and incorporated into an immunochromatographic strip. The developed strip was able to detect as low as 105 CFU/mL of K. pneumoniae. Regarding specificity, it showed negative results with both Escherichia coli and Enterobacter cloacae. More importantly, in a pilot study using neonatal sepsis cases blood specimens, the developed strip selectively gave positive results within 20 min with those infected with K. pneumoniae without prior sample processing. However, it gave negative results in cases infected with other bacterial species.

scholarly journals Global Dissemination ofblaKPCinto Bacterial Species beyond Klebsiella pneumoniae andIn VitroSusceptibility to Ceftazidime-Avibactam and Aztreonam-Avibactam

2016 ◽  
Vol 60 (8) ◽  
pp. 4490-4500 ◽  
Author(s):  
Krystyna M. Kazmierczak ◽  
Douglas J. Biedenbach ◽  
Meredith Hackel ◽  
Sharon Rabine ◽  
Boudewijn L. M. de Jonge ◽  
...  
Keyword(s):  
United States ◽  
Klebsiella Pneumoniae ◽  
Molecular Mechanisms ◽  
Bacterial Species ◽  
The United States ◽  
Asia Pacific ◽  
Gram Negative ◽  
Content Type ◽  
Global Problem

ABSTRACTTheKlebsiella pneumoniaecarbapenemase (KPC), first described in the United States in 1996, is now a widespread global problem in several Gram-negative species. A worldwide surveillance study collected Gram-negative pathogens from 202 global sites in 40 countries during 2012 to 2014 and determined susceptibility to β-lactams and other class agents by broth microdilution testing. Molecular mechanisms of β-lactam resistance among carbapenem-nonsusceptibleEnterobacteriaceaeandPseudomonas aeruginosawere determined using PCR and sequencing. Genes encoding KPC enzymes were found in 586 isolates from 22 countries (76 medical centers), including countries in the Asia-Pacific region (32 isolates), Europe (264 isolates), Latin America (210 isolates), and the Middle East (19 isolates, Israel only) and the United States (61 isolates). The majority of isolates wereK. pneumoniae(83.4%); however, KPC was detected in 13 additional species. KPC-2 (69.6%) was more common than KPC-3 (29.5%), with regional variation observed. A novel KPC variant, KPC-18 (KPC-3[V8I]), was identified during the study. Few antimicrobial agents tested remained effectivein vitroagainst KPC-producing isolates, with ceftazidime-avibactam (MIC90, 4 μg/ml), aztreonam-avibactam (MIC90, 0.5 μg/ml), and tigecycline (MIC90, 2 μg/ml) retaining the greatest activity againstEnterobacteriaceaecocarrying KPC and other β-lactamases, whereas colistin (MIC90, 2 μg/ml) demonstrated the greatestin vitroactivity against KPC-positiveP. aeruginosa. This analysis of surveillance data demonstrated that KPC is widely disseminated. KPC was found in multiple species ofEnterobacteriaceaeandP. aeruginosaand has now become a global problem.

scholarly journals Neurotoxin Gene Profiling of Clostridium botulinum Types C and D Native to Different Countries within Europe

2012 ◽  
Vol 78 (9) ◽  
pp. 3120-3127 ◽  
Author(s):  
Cedric Woudstra ◽  
Hanna Skarin ◽  
Fabrizio Anniballi ◽  
Lucia Fenicia ◽  
Luca Bano ◽  
...  
Keyword(s):  
Clostridium Botulinum ◽  
Limit Of Detection ◽  
Bacterial Species ◽  
Simultaneous Detection ◽  
Geographical Location ◽  
Gene Profiling ◽  
Animal Origin ◽  
Bacterial Strains ◽  
Content Type ◽  
Pcr Assays

ABSTRACTClostridium botulinumtypes C and D, as well as their mosaic variants C-D and D-C, are associated with avian and mammalian botulism. This study reports on the development of low-density macroarrays based on the GeneDisc cycler platform (Pall-GeneDisc Technologies) applied to the simultaneous detection of theC. botulinumsubtypes C, C-D, D, and D-C. The limit of detection of the PCR assays was 38 fg of total DNA, corresponding to 15 genome copies. Artificially contaminated samples of cecum showed a limit of detection below 50 spores/g. The tests were performed with a large variety of bacterial strains, includingC. botulinumtypes C (n= 12), C-D (n= 29), D (n= 5), and D-C (n= 10), other botulinum neurotoxin (BoNT)-producingClostridiumstrains (n= 20), non-BoNT-producing clostridia (n= 20), and other bacterial species (n= 23), and showed a high specificity. These PCR assays were compared to previously published real-time PCRs for the detection ofC. botulinumin 292 samples collected from cases of botulism events in four European regions. The majority of the samples originated from wild birds (n= 108), poultry (n= 60), and bovines (n= 56). Among the 292 samples, 144 were positive for either thebont/C-D or thebont/D-C gene by using the GeneDisc arrays. The reliability of the results tallied to 97.94%. Interestingly, only BoNT mosaics, types C-D and D-C, were found in naturally contaminated samples whatever their animal origin and their geographical location. Further investigations should now be performed in order to check that mosaic types dominate in Europe and that acquisition of mosaic types helps in survival or adaptation to particular niche.

scholarly journals Serum Antibody Responses against Carbapenem-Resistant Klebsiella pneumoniae in Infected Patients

mSphere ◽  
2021 ◽  
Vol 6 (2) ◽  
Author(s):  
Kasturi Banerjee ◽  
Michael P. Motley ◽  
Elizabeth Diago-Navarro ◽  
Bettina C. Fries
Keyword(s):  
Klebsiella Pneumoniae ◽  
Capsular Polysaccharide ◽  
Serum Antibody ◽  
Antibody Responses ◽  
Capsular Polysaccharides ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Reactive Properties

ABSTRACT Capsular polysaccharide (CPS) heterogeneity within carbapenem-resistant Klebsiella pneumoniae (CR-Kp) strain sequence type 258 (ST258) must be considered when developing CPS-based vaccines. Here, we sought to characterize CPS-specific antibody responses elicited by CR-Kp-infected patients. Plasma and bacterial isolates were collected from 33 hospital patients with positive CR-Kp cultures. Isolate capsules were typed by wzi sequencing. Reactivity and measures of efficacy of patient antibodies were studied against 3 prevalent CR-Kp CPS types (wzi29, wzi154, and wzi50). High IgG titers against wzi154 and wzi50 CPS were documented in 79% of infected patients. Patient-derived (PD) IgGs agglutinated CR-Kp and limited growth better than naive IgG and promoted phagocytosis of strains across the serotype isolated from their donors. Additionally, poly-IgG from wzi50 and wzi154 patients promoted phagocytosis of nonconcordant CR-Kp serotypes. Such effects were lost when poly-IgG was depleted of CPS-specific IgG. Additionally, mice infected with wzi50, wzi154, and wzi29 CR-Kp strains preopsonized with wzi50 patient-derived IgG exhibited lower lung CFU than controls. Depletion of wzi50 antibodies (Abs) reversed this effect in wzi50 and wzi154 infections, whereas wzi154 Ab depletion reduced poly-IgG efficacy against wzi29 CR-Kp. We are the first to report cross-reactive properties of CPS-specific Abs from CR-Kp patients through both in vitro and in vivo models. IMPORTANCE Carbapenem-resistant Klebsiella pneumoniae is a rapidly emerging public health threat that can cause fatal infections in up to 50% of affected patients. Due to its resistance to nearly all antimicrobials, development of alternate therapies like antibodies and vaccines is urgently needed. Capsular polysaccharides constitute important targets, as they are crucial for Klebsiella pneumoniae pathogenesis. Capsular polysaccharides are very diverse and, therefore, studying the host’s capsule-type specific antibodies is crucial to develop effective anti-CPS immunotherapies. In this study, we are the first to characterize humoral responses in infected patients against carbapenem-resistant Klebsiella pneumoniae expressing different wzi capsule types. This study is the first to report the efficacy of cross-reactive properties of CPS-specific Abs in both in vitro and in vivo models.

scholarly journals Development of an immunochromatographic lateral flow strip for the simultaneous detection of aminoglycoside residues in milk

RSC Advances ◽  
2018 ◽  
Vol 8 (17) ◽  
pp. 9580-9586 ◽  
Author(s):  
Yaning Sun ◽  
Jifei Yang ◽  
Suzhen Yang ◽  
Qingbo Sang ◽  
Man Teng ◽  
...  
Keyword(s):  
Colloidal Gold ◽  
Simultaneous Detection ◽  
Lateral Flow ◽  
Quantitative Detection ◽  
Lateral Flow Strip ◽  
Immunochromatographic Strip ◽  
Gentamicin Sulfate ◽  
Kanamycin Sulfate ◽  
Neomycin Sulfate

A colloidal gold-based immunochromatographic strip has been developed for the rapid, simultaneous, semi-quantitative and quantitative detection of several aminoglycoside residues in milk, including gentamicin sulfate (GM), neomycin sulfate (NEO) and kanamycin sulfate (KN).

scholarly journals In Vivo Pharmacokinetic and Pharmacodynamic Profiles of Antofloxacin against Klebsiella pneumoniae in a Neutropenic Murine Lung Infection Model

2017 ◽  
Vol 61 (5) ◽  
Author(s):  
Yu-Feng Zhou ◽  
Meng-Ting Tao ◽  
Wei Huo ◽  
Xiao-Ping Liao ◽  
Jian Sun ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Bacterial Species ◽  
Dose Range ◽  
Lung Infection ◽  
Free Drug ◽  
Dose Fractionation ◽  
Infection Model ◽  
Content Type ◽  
Murine Lung ◽  
Tract Infections

ABSTRACT Antofloxacin is a novel broad-spectrum fluoroquinolone under development for the treatment of infections caused by a diverse group of bacterial species. We explored the pharmacodynamic (PD) profile and targets of antofloxacin against seven Klebsiella pneumoniae isolates by using a neutropenic murine lung infection model. Plasma and bronchopulmonary pharmacokinetic (PK) studies were conducted at single subcutaneous doses of 2.5, 10, 40, and 160 mg/kg of body weight. Mice were infected intratracheally with K. pneumoniae and treated using 2-fold-increasing total doses of antofloxacin ranging from 2.5 to 160 mg/kg/24 h administered in 1, 2, 3, or 4 doses. The E max Hill equation was used to model the dose-response data. Antofloxacin could penetrate the lung epithelial lining fluid (ELF) with pharmacokinetics similar to those in plasma with linear elimination half-lives over the dose range. All study strains showed a 3-log10 or greater reduction in bacterial burden and prolonged postantibiotic effects (PAEs) ranging from 3.2 to 5.3 h. Dose fractionation response curves were steep, and the free-drug area under the concentration-time curve over 24 h (AUC0–24)/MIC ratio was the PD index most closely linked to efficacy (R 2 = 0.96). The mean free-drug AUC0–24/MIC ratios required to achieve net bacterial stasis, a 1-log10 kill, and a 2-log10 kill for each isolate were 52.6, 89.9, and 164.9, respectively. When integrated with human PK data, these PD targets could provide a framework for further optimization of dosing regimens. This could make antofloxacin an attractive option for the treatment of respiratory tract infections involving K. pneumoniae.

scholarly journals Multiplex PCR for Identification of Two Capsular Types in Epidemic KPC-Producing Klebsiella pneumoniae Sequence Type 258 Strains

2014 ◽  
Vol 58 (7) ◽  
pp. 4196-4199 ◽  
Author(s):  
Liang Chen ◽  
Kalyan D. Chavda ◽  
Jacqueline Findlay ◽  
Gisele Peirano ◽  
Katie Hopkins ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Multiplex Pcr ◽  
Capsular Polysaccharide ◽  
Sequence Type ◽  
Pcr Assay ◽  
Content Type ◽  
Multiplex Pcr Assay ◽  
Polysaccharide Synthesis ◽  
Sequence Types

ABSTRACTWe developed a multiplex PCR assay capable of identifying two capsular polysaccharide synthesis sequence types (sequence type 258 [ST258]cps-1andcps-2) in epidemicKlebsiella pneumoniaeST258 strains. The assay performed with excellent sensitivity (100%) and specificity (100%) for identifyingcpstypes in 60 ST258K. pneumoniaesequenced isolates. The screening of 419 ST258 clonal isolates revealed a significant association betweencpstype andK. pneumoniaecarbapenemase (KPC) variant:cps-1is largely associated with KPC-2, whilecps-2is primarily associated with KPC-3.

scholarly journals Population Structure of KPC-Producing Klebsiella pneumoniae Isolates from Midwestern U.S. Hospitals

2014 ◽  
Vol 58 (8) ◽  
pp. 4961-4965 ◽  
Author(s):  
Meredith S. Wright ◽  
Federico Perez ◽  
Lauren Brinkac ◽  
Michael R. Jacobs ◽  
Keith Kaye ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Capsular Polysaccharide ◽  
Sequence Type ◽  
Single Nucleotide Variants ◽  
Single Nucleotide ◽  
Content Type ◽  
Strain Diversity ◽  
Carbapenem Resistant ◽  
Genetic Context ◽  
Over Time

ABSTRACTGenome sequencing of carbapenem-resistantKlebsiella pneumoniaeisolates from regional U.S. hospitals was used to characterize strain diversity and theblaKPCgenetic context. A phylogeny based on core single-nucleotide variants (SNVs) supports a division of sequence type 258 (ST258) into two distinct groups. The primary differences between the groups are in the capsular polysaccharide locus (cps) and their plasmid contents. A strict association between clade and KPC variant was found. TheblaKPCgene was found on variants of two plasmid backbones. This study indicates that highly similarK. pneumoniaesubpopulations coexist within the same hospitals over time.

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