scholarly journals Rapid Detection of Methicillin Resistance in Staphylococcus aureus Isolates by the MRSA-Screen Latex Agglutination Test

1999 ◽  
Vol 37 (9) ◽  
pp. 3029-3030 ◽  
Author(s):  
Willem B. van Leeuwen ◽  
Cindy van Pelt ◽  
Ad Luijendijk ◽  
Henri A. Verbrugh ◽  
Wil H. F. Goessens
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Screen Test ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant ◽  
Negative Results ◽  
Specificity And Sensitivity ◽  
Latex Test

The slide agglutination test MRSA-Screen (Denka Seiken Co., Niigata, Japan) was compared with the mecA PCR (“gold standard”) for the detection of methicillin resistance inStaphylococcus aureus. The MRSA-Screen test detected the penicillin-binding protein 2a (PBP2a) antigen in 87 of 90 genetically diverse methicillin-resistant S. aureus (MRSA) stock culture strains, leading to a sensitivity of 97%. The three discrepant MRSA strains displayed positive results only after induction of themecA gene by exposure to methicillin. Both mecAPCR and MRSA-Screen displayed negative results among the methicillin-susceptible S. aureus strains (n = 106), as well as for Micrococcus spp. (n = 10), members of the familyEnterobacteriaceae (n = 10),Streptococcus pneumoniae (n = 10), andEnterococcus spp. (n = 10) (specificity = 100%). Producing the same PBP2a antigen, all 10 methicillin-resistant Staphylococcus epidermidis strains score positived in both the latex test and the mecA PCR. Consequently, the MRSA-Screen test should be applied only after identification of the MRSA strain to the species level to rule out coagulase-negative staphylococci. In conclusion, due to excellent specificity and sensitivity the MRSA-Screen latex test has the potential to be successfully used for routine applications in the microbiology laboratory.

scholarly journals Rapid Slide Latex Agglutination Test for Detection of Methicillin Resistance in Staphylococcus aureus

1999 ◽  
Vol 37 (9) ◽  
pp. 2789-2792 ◽  
Author(s):  
Arjanne van Griethuysen ◽  
Miranda Pouw ◽  
Nan van Leeuwen ◽  
Max Heck ◽  
Piet Willemse ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Screen Test ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Latex Agglutination Test ◽  
Specific Test ◽  
Methicillin Resistant ◽  
Phage Types ◽  
Test Specificity

The MRSA screen test (Denka Seiken Co., Ltd.), a commercially available, rapid (20-min) slide latex agglutination test for the determination of methicillin resistance by detection of PBP 2a inStaphylococcus aureus, was compared with the oxacillin agar screen test and PCR detection of the mecA gene. A total of 563 S. aureus isolates were tested. Two hundred ninety-six of the isolates were methicillin-susceptible isolates from cultures of blood from consecutive patients. Also, 267 methicillin-resistant isolates that comprised 248 different phage types were tested. Methicillin resistance was defined as the presence of themecA gene. Of the 267 mecA gene-positive isolates, 263 were positive by the MRSA screen test (sensitivity, 98.5%), and all the mecA-gene negative strains were negative by the MRSA screen test (specificity, 100%). The oxacillin agar screen test detected methicillin resistance in 250 of themecA gene-positive isolates (sensitivity, 93.6%). The sensitivity of the MRSA screen test was statistically significantly higher than the sensitivity of the oxacillin agar screen test (P < 0.05). The MRSA screen test is a highly sensitive and specific test for the detection of methicillin resistance. Also, it offers results within half an hour and is easy to perform, which makes this test a valuable tool in the ongoing battle against methicillin-resistant S. aureus.

scholarly journals Detection of mecA gene by latex agglutination and its molecular analysis by PCR in methicillin resistant staphylococcus aureus.

2020 ◽  
Vol 27 (07) ◽  
pp. 1363-1370
Author(s):  
Aneela Khawaja ◽  
Iffat Javed ◽  
Sohaila Mushtaq ◽  
Saeed Anwar ◽  
Faiqa Arshad ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Medical Institute ◽  
Cross Sectional ◽  
Methicillin Resistant ◽  
Agglutination Method ◽  
Resistant Strains

Antimicrobial resistance (AMR) is a devastating question that is threatening the health globally. The extensive and indiscriminative use of antibiotics has evolved a notorious resistance in Staphylococcus aureus.  This resistance developed through possession of mecA gene, which codes for modified penicillin binding protein (PBP2a) and the emergent strain being labeled “methicillin resistant Staphylococcus aureus”. Conventional phenotypic techniques for detection of MRSA rely on standardization of cultural characteristics. The drawbacks of diagnostic error to report MRSA include: poor prognosis, expensive treatment, dissemination of multi-drug resistant strains and even treatment failure. Latex agglutination method can be adopted as a more accurate and quick strategy for rapid detection of methicillin resistance. Objectives: To compare detection of mecA gene in methicillin resistant isolates of Staphylococcus aureus by latex agglutination and PCR; by assessing the sensitivity and specificity of both methods. Study Design: Descriptive Cross-Sectional study. Setting: Pathology Department, Post Graduate Medical Institute, Lahore. Period: From January 2015 to December 2015; according to standard operating procedures at Microbiology laboratory. Material & Methods: A total 713 consecutive, non-duplicate isolates of Staphylococcus aureus were processed. Methicillin resistance was determined using cefoxitin (30mg) by Kirby-Bauer method using CLSI guideline (2016), latex agglutination method; and PCR for mecA gene. Results: The results showed that out of 713 Staphylococcus aureus isolates, 92 (12.90%) isolates were resistant to cefoxitin and were labelled as MRSA. majority MRSA isolates recovered from pus (44.57%) and wound swab (20.65%), followed by blood (13.04%), fluid (8.70%), CSF (4.35%), CVP (3.26%), HVS (3.26%) and tracheal secretion (2.17%). By latex agglutination method, 87 (94.50%) were positive for PBP2a; while on PCR mecA gene was detected only in 82 (89.10%) MRSA isolates. When assessed with PCR (gold standard) the sensitivity and diagnostic accuracy of latex agglutination was 100% and 94.57%, respectively. Conclusion: Latex agglutination test can be employed as rapid and reliable diagnostic technique in MRSA isolates for mecA gene detection, where resources for molecular methods are inadequate. This can effectually lessen the misdiagnosis of resistant strains, and over/ ill-use of antibiotics.

scholarly journals Mannitol Fermenting Methicillin-Resistant Coagulase Negative Staphylococci Isolated From Diabetic Foot Infections

2020 ◽  
Vol 11 (4) ◽  
pp. 6095-6101
Author(s):  
Samira Fattah Hamid ◽  
Aza Bahadeen Taha
Keyword(s):  
Staphylococcus Aureus ◽  
Diabetic Foot ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Rrna Gene ◽  
Human Pathogens ◽  
Coagulase Negative Staphylococci ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Diabetic Foot Infections

Detection of mannitol fermenting coagulase-negative staphylococci is frequently unnoticed when Staphylococcus aureus is screening in the laboratory. On the other hand, the emergence of coagulase-negative staphylococci as critical human pathogens need dependable methods for the identification of clinically significant coagulase-negative staphylococci to understand the epidemiology of infections caused by these bacteria. The study aimed to identify mannitol fermenting coagulase-negative staphylococci that assumed to be Staphylococcus aureus as they formed yellow colonies on Mannitol Salt agar plates. Samples were taken from eighty-four patients with diabetic foot infections. The specimen was cultured on Blood agar and Mannitol Salt agar. Mannitol fermenting coagulase-negative staphylococci isolates diagnosed through Vitek2 system then confirmed by detecting 16S rRNA gene and absence of the nuc gene. Antibiotic sensitivity and methicillin resistance were detected by Vitek2 system, then methicillin resistance was confirmed by Oxacillin Salt Agar Screen test and detection of the mecA gene. Out of 81 Staphylococcus isolated from foot and nose of diabetic foot patients, twenty isolates were mannitol fermenting coagulase-negative staphylococci, they related to following species; Staphylococcus haemolyticus, staphylococcus lentus, Staphylococcus xylosus, Staphylococcus lugdunensis, Staphylococcus hominis, Staphylococcus galinarum and Staphylococcus saprophyticus). The majority of them (85%) were phenotypically methicillin-resistant and genotypically harbouring mecA gene. 80% were resistant to Erythromycin, 70% to Clindamycin, 35% to Trimethoprim-Sulphamethoxazole, 30% to Gentamicin and Rifampicin, 15% to Levofloxacin and Teicoplanin. 30% expressed inducible clindamycin resistance.

scholarly journals Evaluation of MRSA-Screen, a Simple Anti-PBP 2a Slide Latex Agglutination Kit, for Rapid Detection of Methicillin Resistance in Staphylococcus aureus

1999 ◽  
Vol 37 (5) ◽  
pp. 1591-1594 ◽  
Author(s):  
Matthias Cavassini ◽  
Aline Wenger ◽  
Katia Jaton ◽  
Dominique S. Blanc ◽  
Jacques Bille
Keyword(s):  
Staphylococcus Aureus ◽  
Monoclonal Antibody ◽  
Screening Test ◽  
Rapid Detection ◽  
Methicillin Resistance ◽  
Screen Test ◽  
Latex Agglutination ◽  
Disk Diffusion ◽  
Diffusion Test ◽  
Disk Diffusion Test

The MRSA-Screen test (Denka Seiken Co., Ltd., Tokyo, Japan), consisting of a slide latex agglutination kit that detects PBP 2a with a monoclonal antibody, was blindly compared to the oxacillin disk diffusion test, the oxacillin-salt agar screen, and PCR of themecA gene for the detection of methicillin resistance inStaphylococcus aureus. A total of 120 methicillin-susceptible S. aureus (MSSA) and 80 methicillin-resistant S. aureus (MRSA) isolates, defined by the absence or presence of the mecA gene, respectively, were tested. The MRSA-Screen test, the oxacillin disk diffusion test, and the oxacillin-salt agar screening test showed sensitivities of 100, 61.3, and 82.5% and specificities of 99.2, 96.7, and 98.3%, respectively. We conclude that the MRSA-Screen is a very accurate, reliable, and fast test (15 min) for differentiation of MRSA from MSSA colonies on agar plates.

scholarly journals Molecular detection and typing of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India

2019 ◽  
Vol 12 (11) ◽  
pp. 1760-1768 ◽  
Author(s):  
Nimita Venugopal ◽  
Susweta Mitra ◽  
Rituparna Tewari ◽  
Feroze Ganaie ◽  
Rajeswari Shome ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Animal Health ◽  
Sccmec Type ◽  
Meca Gene ◽  
Molecular Characteristics ◽  
Rrna Gene ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant

Background and Aim: Methicillin-resistant staphylococci are among the emerging pathogens which have become a threat to both human and animal health. The present investigation intended to examine the occurrence and the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) recovered from cattle, its handlers, and their environment. Materials and Methods: A total of 666 specimens were subjected to culture method and genus-specific polymerase chain reaction (PCR) for the identification of Staphylococcus. Methicillin resistance was substantiated by PCR identification of mecA and mecC resistance determinants. Species-specific identification of mecA positive isolates was conducted by multiplex PCR. The unidentified species were deciphered by 16S rRNA gene sequencing approach. The mecA positive isolates were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). Results: Duplex PCR identified 728 Staphylococcus isolates, of which 66 (9%) were positive for mecA gene. MRSA constituted 24% of the total mecA positive isolates. Among MRCoNS, Staphylococcus epidermidis (42%), and Staphylococcus haemolyticus (11%) were the most common species identified. Overall, 47% of the mecA positive isolates belonged to SCCmec type V. MLST analysis showed eight different sequence types (STs) among MRSA isolates of which five were novel STs. Among methicillin-resistant S. epidermidis, 19 different STs were found, of which nine novel STs were detected. Conclusion: The increase in the prevalence of mecA positive staphylococci, especially MRCoNS in cattle is a great concern in view of their transmission potential. Hence, continuous monitoring and molecular characterization of methicillin-resistant staphylococci should be elucidated in human and animal sectors so as to prevent the spread of these resistant pathogens.

Evaluation of a Latex Agglutination Test for Rapid Identification of Staphylococcus aureus from Foods

1993 ◽  
Vol 56 (9) ◽  
pp. 759-762 ◽  
Author(s):  
TSUNG C. CHANG ◽  
SU H. HUANG
Keyword(s):  
Staphylococcus Aureus ◽  
Immunoglobulin G ◽  
Protein A ◽  
Rapid Identification ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Polystyrene Latex ◽  
Latex Agglutination Test ◽  
Overnight Incubation ◽  
Latex Test

A latex agglutination kit (AUREUS TEST™) for the rapid identification of Staphylococcus aureus was evaluated in this study. The reagent consists of polystyrene latex particles sensitized with rabbit anti-protein A immunoglobulin G and fibrinogen. Due to the respective binding of bacterial protein A with immunoglobulin G and coagulase with fibrinogen, an agglutination reaction occurs within 1 min when the latex is mixed with a suspension of S. aureus. Of 157 S. aureus isolates (138 from foods) and 110 non-S. aureus isolates (58 species belonging to 19 genera), the sensitivity and specificity of the latex test were 100 and 94.4–100%, respectively. The results were comparable to the conventional coagulase test. Therefore, it is proposed that suspicious colonies of S. aureus on Baird-Parker agar medium be subcultured to tryptic soy agar for overnight incubation. Cultures grown on tryptic soy agar are used for latex agglutination test. The latex agglutination test can be finished within minutes and could be used as an alternative rapid procedure of the coagulase test, which needs several hours or even overnight incubation for completion. In addition, all S. aureus isolates grown on Baird-Parker agar also could be correctly identified by the latex reagent.

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