Simple Method for Quantitation of Cell‐Bound Protein A on <i>Staphylococcus aureus</i> Cells by Means of Hemagglutination with Sheep Erythrocytes Differentially Sensitized with Rabbit Antibody and Its Clinical Application

Hwang, Soo Myung; Seki, Kazuhiko; Murai, Miyo; Sakurada, Junji; Nishihara, Shoko; Maeda, Toshiro; Masuda, Shōgo

doi:10.1111/j.1348-0421.1989.tb01509.x

Cited by 5 publications

(4 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…I t is interesting that SpA-bearing strain is rare in bovines of which immunoglobulins have low affinity to SpA (9). Recently, it was reported that over twothirds of S. aureus strains isolated from clinical materials bear no or a trace amount of cell-bound SpA (11). Since S. aureus are equipped with many other mechanisms for the adhesion to host cells, S. aureus might adhere to various host tissues and cells, according to the immunological state of the host and its location in the host by the most suitable binding strategies.…”

Section: Discussionmentioning

confidence: 99%

The Adhesion of Protein A‐Bearing Staphylococcus aureus Organisms to Soluble‐Staphylococcal‐Antigens‐Coated HeLa Cells Mediated by Specific Antibodies

Murai

Sakurada

Seki

et al. 1991

Microbiology and Immunology

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Discussionmentioning

confidence: 99%

The Adhesion of Protein A‐Bearing Staphylococcus aureus Organisms to Soluble‐Staphylococcal‐Antigens‐Coated HeLa Cells Mediated by Specific Antibodies

Murai

Sakurada

Seki

et al. 1991

Microbiology and Immunology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Winblad and Ericson (14) reported the availability of hemagglutination test for the detection of cell-wall-binding SpA of S. aureus and its quantitative version has been devised in our laboratory (4). In the present investigation, the hemagglutination method was applied for the detection of SpA contained in E. coli transformants.…”

Section: Discussionmentioning

confidence: 99%

Some Improved Versions of Methods for the Indirect Detection of Staphylococcal Protein A Gene Expressed in Escherichia coli

Sakurada

Sumi

1992

Microbiology and Immunology

Self Cite

View full text Add to dashboard Cite

Several versions of methods for the indirect detection of expression of staphylococcal protein A gene (spa) in Escherichia coli (E. coli) were devised by making use of biological properties of staphylococcal protein A (SpA). i) Hemagglutination of sheep red blood cells (SRBC) sensitized with anti-SRBC-antibodies using heattreated spa-transformed E. coli organisms; Native spa-transformed E. coli organisms did not agglutinate the sensitized SRBC. The heat-treatment (60 C, 4 hr) of the transformants, however, caused positive hemagglutination like SpA-positive Staphylococcus aureus (S. aureus) organisms. ii) Halo formation around colonies on agar plates containing normal dog serum, which is originally used for the detection of SpA of S. aureus. A mutant strain NMJ was isolated, which showed formation of the halo of precipitate due to interaction between immunoglobulin and SpA. iii) A new version of immunodetection; After lysis of the transformants grown on a nitrocellulose membrane by alkali, SpA could be directly detected by immuno-detection procedures after inactivation of endogenous peroxidase in bacteria by phenylhydrazine and hydrogen peroxide.Usually, the proteins synthesized by exogenous genes introduced in coli (E. coli) are not secreted out of the bacterial cell, even if the genes are properly expressed. This property makes the screening of a target gene-containing cell cumbersome, even if the proteins themselves can be detected by simple methods.There have been few simple methods detecting products synthesized by exogenous genes as the general gene-screening. The staphylococcal protein A (SpA) molecules produced by staphylococcal protein A gene (spa)-transformed E. coli are not secreted either (5). So far, some reports on the cloning of spa have been published; the screening of spa in all of them was accomplished by the ordinary methods without making use of the properties of SpA and bacteria (2, 5). The present study is an attempt to provide versatile and convenient methods for detection of transformed genes using SpA as a model for other genes.

show abstract

“…The Cowan strain which is known to be a good protein A producer was used as a positive control (5). The titer of protein A was expressed as 2 <nl) where n=well number showing haemagglutination.…”

Section: Protein a Titresmentioning

confidence: 99%

Biological characteristics of methicillin resistant <em>Staphylococcus aureus</em> strains isolated from hospitals in Colombo

Silva

Mendis

1996

Cey. J. Med. Sci.

View full text Add to dashboard Cite

This study was done to determine the antibiogram and biological characteristics of Methicillin resistant Staphylococcus aureus (MRSA) clinical isolates, to detect the existence of a multi-resistant strain conforming to an epidemic MRSA strain.Specimens received from some wards of the General Hospital Colombo, and other hospitals in Colombo and from a few out patients, during a fourteen month period, yielded sixty five MRSA strains. These strains were characterized by the following tests: Antibiotic susceptibility tests, coagulase production with human and rabbit plasma, urea hydrolysis at 24 and 48 hours, protein A levels, and pigment production.Forty five MRSA strains were resistant to the same seven antibiotics in addition to methicillin, and were arbitrarily named group A. This was the predominant strain in the surgical units (SU), plastic surgery unit (PSU) and burns unit (BU).Ninety seven percent of MRSA were resistant to at least one aminoglycoside tested. Resistance to fusidic acid (8%), clindamicin (8%) and rifampicin (4.6%) was rare in MRSA's. All MRSA strains tested were sensitive to vancomycin.The multi-resistant strain -group A, differed from the miscellaneous strains (group B) in the following way; they were marginally stronger in the coagulase test than group B, urea hydrolysis was greater at 24 and 48 hours (P=0.0125 and P=0.0165), protein A levels were marginally higher and the strains were more pigmented. Protein A levels were however low in all the MRSA's when compared to the Cowan strain of S. aureus. The multi-resistant, group A strain to some extent showed the biological characteristics of an epidemic strain of MRSA (EMRSA) described in the literature, which have spread among hospitalised patients and caused serious outbreaks of hospital infections. It remains to be seen if this strain too, will be the likely cause of outbreaks of serious hospital infections in Colombo, as has been described elsewhere in the world.

show abstract

Simple Method for Quantitation of Cell‐Bound Protein A on Staphylococcus aureus Cells by Means of Hemagglutination with Sheep Erythrocytes Differentially Sensitized with Rabbit Antibody and Its Clinical Application

Cited by 5 publications

References 20 publications

The Adhesion of Protein A‐Bearing Staphylococcus aureus Organisms to Soluble‐Staphylococcal‐Antigens‐Coated HeLa Cells Mediated by Specific Antibodies

The Adhesion of Protein A‐Bearing Staphylococcus aureus Organisms to Soluble‐Staphylococcal‐Antigens‐Coated HeLa Cells Mediated by Specific Antibodies

Some Improved Versions of Methods for the Indirect Detection of Staphylococcal Protein A Gene Expressed in Escherichia coli

Biological characteristics of methicillin resistant <em>Staphylococcus aureus</em> strains isolated from hospitals in Colombo

Contact Info

Product

Resources

About