A Rapid Latex Agglutination Assay for the Detection of Penicillin‐Binding Protein 2′

Nakatomi, Yasuo; Sugiyama, Junichi

doi:10.1111/j.1348-0421.1998.tb02347.x

Cited by 79 publications

(41 citation statements)

References 16 publications

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“…For indirect immunoperoxidase staining, anti-Escherichia coli rabbit polyclonal antibody (diluted at 1 : 100; DAKO, Kyoto, Japan), three types of anti-Staphylococcus aureus rabbit polyclonal antibodies (diluted at 1 : 2,560; CH-91, Wood-46, and 3-9D, courtesy of Dr. S. Takeuchi, Fukui Prefectural University), 3 and four types of anti-PBP2Ј mouse monoclonal antibodies (diluted at 1 : 50; B345M, A376M, E303M, and A465M, Denka Seiken, Tokyo, Japan) 6 were used as primary antibodies. The reactivity of the anti-S. aureus antibodies with S. aureus 3 and S. intermedius was confirmed by positive reactions with bacteria smears.…”

Section: Methodsmentioning

confidence: 99%

Abscess-Forming Inflammatory Granulation Tissue with Gram- Positive Cocci and Prominent Eosinophil Infiltration in Cats: Possible Infection of Methicillin-resistant Staphylococcus

Ozaki

Yamagami²,

Nomura³

et al. 2003

Vet Pathol

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Abstract. We occasionally encounter feline cervical or mesenteric lesions diagnosed histopathologically as abscess or inflammatory granulation tissue with eosinophil infiltration. Gram-positive cocci accompany the lesions. In the present study, such lesions obtained from 27 cats were examined to evaluate the histopathologic features and the nature of the causative bacteria. The average age was 7.3 Ϯ 3.5 years. No sex predilection was observed. Most frequent locations of the lesions included the abdominal cavity with/without mesenteric lymph nodes (11/ 27, 41%) and subcutaneous tissue or lymph nodes of the neck (9/27, 33%). Common clinical presentation was a localized mass. Grossly, the lesions contained abscesses in the center and were surrounded by fibrous tissue. Microscopically, the necrotic zone contained bacterial colonies. Large numbers of eosinophils and macrophages infiltrated the area surrounding the necrotic tissue. The surrounding connective fiber-rich granulation tissue demarcated the eosinophilic abscess. The bacteria were Gram-positive cocci in 23 of the 27 cats and were positive for anti-staphylococcus antiserum in 19 of the 23 cats. In 15 out of 17 lesions, the colonies expressed immunoreactivity to penicillin-binding protein 2Ј, which is a drug-resistance gene product of methicillin-resistant Staphylococcus (MRS) species. These findings suggest strongly that MRS causes this type of infectious lesion.

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Section: Methodsmentioning

confidence: 99%

Abscess-Forming Inflammatory Granulation Tissue with Gram- Positive Cocci and Prominent Eosinophil Infiltration in Cats: Possible Infection of Methicillin-resistant Staphylococcus

Ozaki

Yamagami²,

Nomura³

et al. 2003

Vet Pathol

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“…Numerous approaches that improve turnaround time for the identification of MRSA have been described. Fluorescence tests (14), PCR assays (4), or penicillin-binding protein 2a (PBP2a) antibody agglutination tests have been described elsewhere (12). Yet, these require subculture on solid media, and many are unable to determine species and methicillin susceptibility at the same time.…”

mentioning

confidence: 99%

Rapid PCR-Based Identification of Methicillin-Resistant Staphylococcus aureus from Screening Swabs

Jonas¹,

Speck²,

Daschner³

et al. 2002

J Clin Microbiol

153

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A PCR identification of methicillin-resistant Staphylococcus aureus (MRSA), obviating the need for subculture on agar media, was investigated. The combination of MRSA detection by mecA femB PCR with prior enrichment in selective broth was tested for 439 swabs. PCR identified 36 MRSA-positive samples, in concordance with conventional methods.Accurate and rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) in clinical specimens is essential for timely decisions on isolation procedures and effective antimicrobial chemotherapy. Numerous approaches that improve turnaround time for the identification of MRSA have been described. Fluorescence tests (14), PCR assays (4), or penicillin-binding protein 2a (PBP2a) antibody agglutination tests have been described elsewhere (12). Yet, these require subculture on solid media, and many are unable to determine species and methicillin susceptibility at the same time. A simultaneous test of methicillin resistance and species confirmation by a mecA femB duplex PCR has been proposed elsewhere (16). The mecA gene encodes the extra PBP2a, which is unique to methicillin-resistant staphylococci. The femB gene codes for an enzyme important in cross-linking peptidoglycan in various different Staphylococcus spp. The specificity of the femB PCR primers used for DNA amplification of the species S. aureus has been demonstrated previously (8).This study describes the performance of this technique in a clinical setting of moderate MRSA endemicity where large numbers of screens need to be processed on a daily basis. Moreover, the robustness of the test was investigated by determining the number of false-positive readings due to coamplification of femB and mecA from methicillin-susceptible S. aureus (MSSA) and methicillin-resistant coagulase-negative staphylococci (R-CNS) coexisting at the sample site (C. M.

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“…Laboratory standards such as CLSI recommends an additional screening test based on spot inoculation onto NaCl and oxacillincontaining screening plates with a high inoculum (CLSI, 2005). A rapid phenotype identification of MRSA starting from a culture plate can also be performed by a latexagglutination test based on monoclonal antibodies against PBP2a (Nakatoni, Y. and Sugiyama, J., 1998). Assays based on cefoxitin are particularly important for detection of low level oxacillin resistant MRSA (Witte, W., Pasemann, B. et al, 2007).…”

Section: Methods For Antimicrobial Susceptibility Testing (Ast) and Cmentioning

confidence: 99%

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2009

EFSA Journal

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SUMMARYThe European Food Safety Authority (EFSA) asked its Panel on Biological Hazards to deliver a scientific opinion on: The assessment of the Public Health significance of meticillin resistant Staphylococcus aureus (MRSA).There are different states of interaction between S. aureus and its host: infections, carriage or colonisation, and contamination. Meticillin resistant S. aureus (MRSA) can be persistently or intermittently carried by healthy humans, and colonisation is the major risk factor for infection. Infection can be mild to severe and, in some instances, fatal. MRSA are now a major cause of hospital acquired infection in many European countries, with large differences in prevalence and control policies. A limited number of lineages of MRSA tend to predominate in specific geographical locations. CC398 is the MRSA lineage most often associated with asymptomatic carriage in intensively reared food-producing animals. MRSA commonly carry enterotoxin genes but there has been only one report of food intoxication due to MRSA. On the question on what is the risk to human health posed by MRSA associated with foodproducing animals, the Panel concluded that:Livestock-associated MRSA (LA-MRSA) represent only a small proportion of the total number of reports of MRSA infections in the EU. However, this proportion differs between Member States. In some countries with low prevalence of human MRSA infection, CC398 is a major contributor to the overall MRSA burden. In countries with high overall human MRSA prevalence, CC398 is considered of less significance for the public health. CC398 has, albeit rarely, been associated with deep-seated infections of skin and soft tissue, pneumonia and septicaemia in humans. Where CC398 prevalence is high in food-producing animals, people in contact with these live animals (especially farmers and veterinarians, and their families) are at greater risk of colonisation and infection than the general population. The risk to human health from different levels (dose response) of MRSA during carriage in animals (and in the environment) is not known. On the question of what is the importance of food, food-producing animals, and companion animals in the risk of human infection and/or food-borne disease caused by MRSA in both the community and hospital settings, the Panel concluded that:Food may be contaminated by MRSA (including CC398): eating and handling contaminated food is a potential vehicle for transmission. There is currently no evidence for increased risk of human colonisation or infection following contact or consumption of food contaminated by CC398 both in the community and in hospital. MRSA (including CC398) can enter the slaughterhouse in or on animals and occurs on raw meat. Although it may become part of the endemic flora of the slaughterhouse, the risk of infection to slaughterhouse workers and persons handling meat appears to be low based on currently available data.Where CC398 prevalence is high in food-producing animals, people in direct contact with these live animals (espe...

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A Rapid Latex Agglutination Assay for the Detection of Penicillin‐Binding Protein 2′

Cited by 79 publications

References 16 publications

Abscess-Forming Inflammatory Granulation Tissue with Gram- Positive Cocci and Prominent Eosinophil Infiltration in Cats: Possible Infection of Methicillin-resistant Staphylococcus

Abscess-Forming Inflammatory Granulation Tissue with Gram- Positive Cocci and Prominent Eosinophil Infiltration in Cats: Possible Infection of Methicillin-resistant Staphylococcus

Rapid PCR-Based Identification of Methicillin-Resistant Staphylococcus aureus from Screening Swabs

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