A new serotyping method for Klebsiella species: evaluation of the technique.

Riser, Eve; Noone, Paul; Bonnet, Marie-Louise

doi:10.1136/jcp.29.4.305

Cited by 19 publications

(13 citation statements)

References 3 publications

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“…Previous studies have shown that different antisera of K antigens may cross-react with other serotypes (9,13). Dilutions were chosen to improve and distinguish the specificity of the antiserum with strong positive results (13). In the present study, we optimized the working concentration of our antiserum and only false positives with very faint bands for K17, K40, and K69 were observed after 6 min.…”

Section: Resultsmentioning

confidence: 99%

“…Inconspicuous or faint bands were observed at the K1 and/or K2 test lines with serotypes K17, K40, and K69 only after 6 min had elapsed from sample loading. Previous studies have shown that different antisera of K antigens may cross-react with other serotypes (9,13). Dilutions were chosen to improve and distinguish the specificity of the antiserum with strong positive results (13).…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2016

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eIn this study, a novel colloidal gold-based immunochromatographic strip (ICS) containing anti-Klebsiella pneumoniae capsular polysaccharide polyclonal antibodies was developed to specifically detect K. pneumoniae serotypes 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, nontypeable K. pneumoniae strains, and other bacterial species were then used to assess the sensitivity and specificity of these test strips. The detection limit was found to be 10 5 CFU, 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 of K. pneumoniae serotypes K1 and K2.K lebsiella pneumoniae is a Gram-negative, rod-shaped bacterium that is found in the normal flora of the mouth, skin, and intestines. Recently, a new type of invasive infection caused by K. pneumoniae has emerged as a global disease (1). There are 77 different serotypes of the capsular polysaccharides (CPSs) of K. pneumoniae (2), with K2 and K21 being the most prevalent in western countries (3). Recently, by genotyping capsular polysaccharide genes, 79 capsular types were distinguished (4, 5). In contrast, the K1 and K2 serotypes are most often associated with bacteremia, liver abscess, and community-acquired pneumonia in Asia (3). However, reports of this new invasive syndrome have recently been on the rise in western countries (6, 7). In addition to causing human disease, K. pneumoniae serotypes K1 and K2 have also been detected in contaminated animal milk (8). Capsular serotypes have typically been identified by using either a capsular swelling test or countercurrent immunoelectrophoresis (3, 9). However, these conventional methods are time-consuming, require extensive materials and manpower, and often produce falsepositive or false-negative results that are caused by cross-reactions. Another option is PCR, but this analysis requires a long time for preparation, expensive equipment, and skilled technicians, and previous studies have recommended that rapid detection is ideal for early clinical diagnosis and treatment (1).To address these concerns, this study involved the development of a colloidal gold-based immunochromatographic strip (ICS) that contained anti-K. pneumoniae capsular polysaccharide polyclonal antibodies (pAbs) and was able to detect K. pneumoniae serotypes K1 and K2. This testing kit provides a sensitive, rapid, and simultaneous method to detect K. pneumoniae serotypes K1 and K2. MATERIALS AND METHODSBacterial strains. The various serotypes of K. pneumoniae that were used here were obtained from our previous studies in Taiwan and at the Serum Institute in Denmark. The panel of bacterial strains contained al...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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

et al. 2016

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“…Serotyping of capsular antigen (K) has been the classical epidemiologic typing method. Seventy-seven K serotypes have been observed; the serotypes can be determined by a variety of methods (17,18,22). Although serotypes are stable markers and serotyping is reproducible, it has the major limitation of requiring reagents that are not commercially available.…”

Section: Discussionmentioning

confidence: 99%

Molecular epidemiology of plasmid spread among extended broad-spectrum beta-lactamase-producing Klebsiella pneumoniae isolates in a pediatric hospital

et al. 1993

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Over a 12-month period, 43 children in eight different wards of our hospital (Hopital Robert Debre) were infected or colonized with Kiebsiella pneumoniae strains producing extended broad-spectrum 13-lactamases. The epidemiology of the outbreak was studied by a molecular approach including the determination of the 13-lactamase physicochemical parameters and plasmid profiles, as well as analysis of the restriction fragment length polymorphisms of the rDNA regions (ribotyping). The last approach produced 12 and 5 different patterns with EcoRI and HindIH, respectively, thus identifying 15 different ribotypes among the 43 clinical K. pneumoniae strains. However, 60%o of the strains in six wards belonged to only two ribotypes, whereas nine ribotypes were observed only once. Twelve isolates from different wards that were representative of the eight most common ribotypes showed four different 13-lactamase isoelectric focusing patterns and seven different * Corresponding author.

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“…Capsular serotyping has been documented as the most reliable typing technique. Different methods have been used for capsular serotyping, such as capsular swelling (Casewell, 1972), indirect immunofluorescence (Riser, Noone & Poulton, 1976;Riser, Noone & Bonnet, 1976), counter-current immunoelectrophoresis (Palfreyman, 1978) and coagglutination and latex agglutination (Onokodi & Wauters, 1981). These techniques, ofwhich capsular swelling is the most traditional, have been successfully used by several workers for epidemiological purposes (Casewell & Philips, 1978;Riser, Noone & Thompson, 1978;Seal et al 1981;Smith, Digori & Eng, 1982).…”

Section: Introductionmentioning

confidence: 99%

Biochemical and serological investigations on clinical isolates of klebsiella

Simoons-Smit¹,

Vught²,

Kanis³

et al. 1985

J. Hyg.

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SUMMARYA series of 925 clinical isolates of klebsiella was examined by serological and biochemical typing. To perform serological typing (capsular swelling) 77 capsular antisera were prepared, tested against the type strains and grouped in 13 pools. With this serotyping method 800 ofthe cultures were typable and 63 distinct types could be recognized.All strains were typable biochemically by means of the numerical coding system of the API-20E system supplemented by digits derived from 15 additional conventional biochemical tests. With the API-20E system 24 different biotypes could be distinguished whereas the combination of API-20E and the 15 additional tests produced 93 biotypes. Maximum discrimination of strains was achieved by the combination of serological and biochemical typing (256 bioserotypes). The reproducibility, typability and discriminating power of the biotyping system was not inferior to serotyping. For epidemiological purposes biotyping can replace serotyping of Klebsiella species, especially in laboratories less well equipped.

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A new serotyping method for Klebsiella species: evaluation of the technique.

Abstract: (Casewell, 1975;Edwards and Ewing, 1968

Cited by 19 publications

References 3 publications

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

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

Molecular epidemiology of plasmid spread among extended broad-spectrum beta-lactamase-producing Klebsiella pneumoniae isolates in a pediatric hospital

Biochemical and serological investigations on clinical isolates of klebsiella

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