Evaluation of MRSA-Screen, a simple anti-PBP 2a slide latex agglutination kit, for rapid detection of methicillin resistance in Staphylococcus aureus

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 the mecA gene for the detection of methicillin resistance in Staphylococcus 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.     

Evaluation of a commercial latex agglutination test for identification of Staphylococcus aureus.

A new, commercially available latex agglutination test (SeroSTAT Staph; Scott Laboratories, Inc., Fiskeville, R.I.) was compared with the tube coagulase and slide coagulase tests as means for identifying Staphylococcus aureus. Of 160 clinical isolates of S. aureus, 159 (99.4%) yielded positive results with the latex agglutination test. Negative latex agglutination test results were obtained with 266 of 267 clinical isolates of Micrococcus spp. and staphylococcal species other than S. aureus (99.6%). The latex agglutination test was found to be a rapid, technically nondemanding method for identifying S. aureus. It was as accurate as the tube coagulase test and more accurate than the slide coagulase test.     

Evaluation of the latex slide agglutination test for identification of Staphylococcus aureus.

This study evaluated the reliability of the latex slide agglutination test for identifying Staphylococcus aureus. A total of 806 clinical isolates of staphylococci were tested for latex agglutination, clumping factor, and free coagulase. Positive latex tests occurred in 98.3% of coagulase-positive strains, whereas 99.6% of coagulase-negative strains gave negative latex tests. It is concluded that in most instances, the latex slide agglutination test is a reliable method for identifying S. aureus in the clinical laboratory.     

Comparison of a yellow latex reagent with other agglutination methods for the identification of Staphylococcus aureus.

A new commercial yellow latex agglutination reagent (Bacto-Staph) was compared with the slide and tube coagulase tests and three other commercial reagents for the identification of 283 Staphylococcus aureus and 54 non-S. aureus staphylococcal strains. Test sensitivities for the identification of S. aureus were as follows: tube coagulase, 99.6%; slide coagulase, 98.6%; Bacto-Staph, 99.6%; Staphylatex, 98.6%; Sero STAT Staph, 98.2%; and Staphyloslide, 97.5%. No false-positive reactions were observed with any of the commercial reagents.     

Rapid and reliable identification of Staphylococcus aureus by a latex agglutination test

A latex slide agglutination test detecting clumping factor and protein A simultaneously is recommended for rapid and reliable routine identification of Staphylococcus aureus. Strains (836) of staphylococci isolated from clinical specimens were examined, all S. aureus strains identified by conventional methods were correctly differentiated by the latex test, and no false-positive results occurred with other staphylococci. The reagent is easy to prepare since plasma is the coating material.     

Evaluation of RapiDEC Staph for identification of Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus saprophyticus.

RapiDEC Staph is a test for presumptive identification of the principal human staphylococcal species, Staphylococcus aureus, S. epidermidis, and S. saprophyticus. The test includes control and test cupules for fluorogenic detection of coagulase and chromogenic substrates for alkaline phosphatase and beta-galactosidase. These tests identify S. aureus, S. epidermidis, and S. saprophyticus, respectively. Positive results with both chromogenic substrates provide a presumptive identification of S. xylosus or S. intermedius (S. xylosus-S. intermedius). Test cupules are inoculated with an organism suspension, and reactions are read after a 2-h incubation. RapiDEC-Staph was evaluated with 303 clinical and stock staphylococcal strains. Identifications were compared with those obtained by the tube coagulase test, a latex slide coagulase test (StaphAUREX), another commercial identification system (Staph-TRAC), and additional conventional tests. RapiDEC-Staph correctly identified 100% of 130 S. aureus strains, 70.3% of 74 S. epidermidis strains, and 81.3% of 32 S. saprophyticus strains. Four of five S. xylosus isolates were called S. xylosus-S. intermedius. Unidentified S. epidermidis and S. saprophyticus strains were called "Staphylococcus spp." Among the 62 other coagulase-negative staphylococci, 4 were misidentified as S. epidermidis and 7 were misidentified as S. saprophyticus. While the sensitivity and specificity of the fluorogenic coagulase test for S. aureus were 100%, failure to detect alkaline phosphatase activity in several S. epidermidis isolates resulted in fewer correct identifications by the RapiDEC-Staph test for this species.     

Evaluation of CHROMagar Staph. aureus, a new chromogenic medium, for isolation and presumptive identification of Staphylococcus aureus from human clinical specimens

CHROMagar Staph. aureus (CSA) is a new chromogenic medium for presumptive identification of Staphylococcus aureus as mauve colonies after 24 h of incubation. We conducted a preliminary study with 100 S. aureus and 45 coagulase-negative Staphylococcus (CoNS) stock isolates plated on CSA. All S. aureus isolates yielded mauve colonies after 24 h of incubation at 37 degrees C, while CoNS isolates grew as blue, white, or beige colonies. Culture on CSA was then prospectively compared to a conventional laboratory method, i.e. , culture on 5% horse blood agar (HBA), catalase test, and latex agglutination test (HBA-catalase-latex), for isolation and presumptive identification of S. aureus from 2,000 consecutive clinical samples. Among the 310 S. aureus isolates recovered by at least one of the two methods, 296 grew as typical mauve colonies on CSA, while only 254 yielded catalase-positive, latex-positive colonies on HBA. The sensitivity of CSA was significantly higher than that of the conventional method (95.5 and 81.9%, respectively; P < 0.001) and allowed the recovery of important clinical isolates that were undetected on blood agar. The specificities of the two methods were not significantly different, although that of CSA was slightly higher (99.4% versus 98.9% for HBA-catalase-latex; P = 0. 08). On the basis of its excellent sensitivity and specificity, ease of identification of positive colonies, and absence of complementary testing, CSA can be recommended as a routine plating medium for presumptive identification of S. aureus in clinical specimens.     

International multicenter evaluation of latex agglutination tests for identification of Staphylococcus aureus

A newly marketed rapid agglutination kit for the identification of Staphylococcus aureus, Slidex Staph Plus (bioMérieux), was compared to Staphaurex Plus (Murex Diagnostics) and Pastorex Staph-Plus (Sanofi Diagnostics Pasteur). The study took place in three clinical microbiology laboratories in three different European countries. A total of 892 staphylococcal isolates, including 278 methicillin-sensitive S. aureus (MSSA) isolates, 171 methicillin-resistant S. aureus (MRSA) isolates, and 443 coagulase-negative staphylococcal isolates, were analyzed. The sensitivities (MSSA/MRSA) and specificities, respectively, were 98. 2% (98.9%/97.1%) and 98.9% for Slidex Staph Plus, 98.2% (98.2%/98. 2%) and 96.2% for Staphaurex Plus, and 98.7% (98.6%/98.8%) and 95.7% for Pastorex Staph Plus. The specificity of the Slidex Staph Plus kit was statistically significantly higher than the specificities of Staphaurex Plus and Pastorex Staph-Plus. The Slidex Staph Plus is a very reliable test for the identification of S. aureus.     

Evaluation of new streptococcal latex grouping kit.

AIMS: To evaluate a new streptococcal latex grouping kit (Shield Diagnostics Ltd) and compare it against an established latex agglutination method (Streptex; Wellcome Diagnostics). METHODS: Two hundred and forty seven strains of streptococci and enterococci were tested with each kit by one operator and according to the manufacturer's instructions. Strains failing to group or giving discordant results were identified to species level. RESULTS: Two discrepant grouping results were observed and 13 non-beta haemolytic streptococci failed to group with either product. The Shield kit successfully identified 232 isolates at 15 minutes of enzyme extraction incubation compared with 224 and 233 on short (15 minutes) and long (1 hour) incubations, respectively, for Streptex (p > 0.23 for both comparisons). On short incubation only, the Shield kit detected significantly more strains of Enterococcus faecium (p = 0.007). The reaction strengths were similar for both kits (p > 0.16). No cross-reactions were observed but the Streptex kit produced significantly fewer tests with visible granularity (p < 0.003). CONCLUSIONS: Although the Shield product appeared to detect group D antigen more readily, overall no important differences in performance were observed. Prospective users of the new method should first become familiar with its characteristics.     

Evaluation of penicillin binding protein 2a latex agglutination assay for identification of methicillin-resistant Staphylococcus aureus directly from blood cultures

The penicillin binding protein 2a (PBP2a) latex agglutination test using a blood culture pellet was compared to the oxacillin screen agar method using isolated colonies. For blood cultures positive for Staphylococcus aureus (n = 70), the direct PBP2a test was 18% sensitive and 100% specific. The PBP2a test shows poor sensitivity when used directly with positive blood cultures.     

Evaluation of new agglutination test for identification of oxacillin-susceptible and oxacillin-resistant Staphylococcus aureus.

A new agglutination test (Monostaph +; Bionor, Skien, Norway) has been developed. This new agglutination test has been compared with two other agglutination tests for the identification of 128 isolates of Staphylococcus aureus and 82 coagulase-negative staphylococci. The sensitivities of both Monostaph + and Pastorex Staph-Plus were excellent (98.7 and 97.4%, respectively) in detection of oxacillin-resistant Staphylococcus aureus. The specificity was 96.4% (two Staphylococcus epidermidis isolates and one Staphylococcus hominis isolate were false positive).     

Reliability of latex agglutination tests for identification of Staphylococcus aureus resistant to oxacillin.

Commercial latex agglutination tests (LATs) for the simultaneous detection of clumping factor and protein A are gaining increased acceptance as a means of identifying Staphylococcus aureus. We evaluated two LATs (Accu-Staph; Carr-Scarborough, Decatur, Ga.; Staphaurex; Wellcome, Dartford, England) with particular emphasis on their ability to correctly identify oxacillin-resistant S. aureus. We tested 59 oxacillin-resistant S. aureus, 136 oxacillin-susceptible S. aureus, and 92 coagulase-negative staphylococcal strains with the two LATs and with thermonuclease, slide clumping factor, tube coagulase, and protein A hemagglutination tests. Clumping factor and protein A were present in 96.9 and 82.1% of our S. aureus strains, respectively. Accu-Staph correctly identified 92.8% and Staphaurex correctly identified 91.3% of S. aureus strains. No significant difference in LAT positivity rates, presence of clumping factor, or presence of protein A was found between oxacillin-resistant and -susceptible S. aureus. Overall, there were 31 false-negative LATs for 20 S. aureus strains, 14 with Accu-Staph and 17 with Staphaurex. Ninety-five percent of these strains possessed either clumping factor or protein A or both when these factors were determined independently. There were five false-positive LATs for four strains of coagulase-negative staphylococci (three Staphylococcus epidermidis and one Staphylococcus warneri), four with Accu-Staph and one with Staphaurex. Clumping factor was present in one S. warneri strain. Thus, the specificities of Accu-Staph, Staphaurex, and the clumping factor test were 95.6, 98.9, and 98.9%, respectively. Our results indicated that LATs identify oxacillin-resistant and -susceptible S. aureus equally well; however, they offer no greater sensitivity or specificity than the clumping factor test for identification of S. aureus.     

Direct identification of Staphylococcus aureus in blood culture fluid with a commercial latex agglutination test.

A commercial latex agglutination slide test (SeroSTAT Staph, Scott Laboratories, Inc., Fiskeville, R.I.) accurately identified Staphylococcus aureus when applied directly to blood culture fluid containing staphylococci. This latex agglutination test exhibited 100% accuracy when 30 seeded aerobic and anaerobic radiometric blood cultures (15 strains of S. aureus, 15 strains of other staphylococcal species) were tested blindly. In 36 actual clinical specimens yielding 16 isolates of S. aureus and 20 isolates of Staphylococcus epidermidis, 94.4% accuracy was achieved. The latex agglutination test provided positive test results before objective criteria of blood culture positivity such as radiometric growth indices and Gram stains became positive.     

Novel latex agglutination method with chicken anti-protein A for detection of Staphylococcus aureus infections.

A latex agglutination assay for the detection of protein A-secreting Staphylococcus aureus strains or strains with protein A in the cell wall is described. The assay utilizes latex particles coated with chicken anti-protein A antibodies. Chicken antibodies do not react with protein G-producing streptococci or rheumatoid factor, thus avoiding false-positive reactions.     

Evaluation of a modified nitrous acid extraction latex agglutination kit for grouping beta-hemolytic streptococci and enterococci.

The rapid and accurate identification of the Lancefield group of beta-hemolytic streptococci and enterococci is an important procedure in clinical laboratories. Latex agglutination techniques are more rapid and technically less demanding than traditional extraction-precipitation methods. Prolex (Pro-Lab Diagnostics, Richmond Hill, Ontario, Canada) is a latex agglutination kit which contains modified nitrous acid reagents to extract antigens and can be used to detect group D antigen in streptococci and enterococci, as well as group A, B, C, F, and G antigens. A total of 302 strains of streptococci and enterococci were tested with this kit. All streptococci of groups A (41 strains), B (39 strains), C (35 strains), D (3 strains), F (10 strains), and G (48 strains) were correctly grouped, as were 125 (97%) of 129 strains of enterococci. Prolex is a reliable method for grouping the beta-hemolytic streptococci and enterococci most frequently encountered in clinical laboratories.     

Evaluation of the methicillin-resistant Staphylococcus aureus (MRSA)-Screen latex agglutination test for detection of MRSA of animal origin

Methicillin (oxacillin)-resistant staphylococci (MRS) have emerged as major clinical and epidemiological pathogens, and there have been frequent reports of MRS infections in the veterinary field. The MRSA-Screen latex agglutination test (Denka Seiken Co., Ltd., Tokyo, Japan) was compared with an oxacillin agar screen test, MIC determination, and mecA PCR assay, the "gold standard." In an analysis of 15 mecA-positive and 48 mecA-negative S. aureus animal isolates, as well as 9 mecA-positive and 147 mecA-negative, coagulase-negative staphylococcal animal isolates, the latex agglutination test surpassed the widely used oxacillin agar screen method and MIC determination, with a sensitivity and a specificity of 100%. The MRSA-Screen test is a reliable and rapid method of detecting MRS in the veterinary field.     

Comparative performances of six agglutination kits assessed by using typical and atypical strains of Staphylococcus aureus.

Six commercial agglutination tests designed for the identification of Staphylococcus aureus were compared by using a strain collection which included 512 staphylococci representing 33 species (318 isolates of Staphylococcus aureus [including 144 oxacillin resistant], 46 S. epidermidis isolates, 15 S. haemolyticus isolates, 12 S. saprophyticus isolates, 29 S. schleiferi isolates, 30 S. lugdunensis isolates, and 62 other coagulase-negative staphylococci). This group also included a proportion of strains with unusual phenotypes (e.g., 19 coagulase-negative S. aureus isolates, 26 clumping factor-negative S. aureus isolates, and 4 S. aureus isolates each with a double deficiency). The overall sensitivity for identification of typical and atypical S. aureus was high with the Staphaurex Plus test (Murex Biotech) (99.7%), the Pastorex Staph Plus test (Sanofi Diagnostics Pasteur) (99.7%), and the Slidex Staph Plus test (bioMérieux) (100%). The overall rate of specificity was affected by the unusual inclusion in this study of a high proportion of non-S. aureus species, such as S. lugdunensis and S. schleiferi, which express a clumping factor and therefore produce a positive result with the agglutination tests.     

Evaluation of a new slide latex agglutination test for diagnosis of vaginal candidosis

A new commercial slide latex particle agglutination test for rapid (2 min) diagnosis of vaginal candidosis was evaluated and compared with conventional methods. Of the 263 women studied, 63 (23.9%) had yeasts in the vagina. Clinical signs of vulvitis or vaginitis were seen in 23 women (8.8%) and 40(15.2%) were harbouring yeasts without clinical signs. Yeast counts were generally higher in women with clinical signs of vaginal candidosis than in those without. The test was positive in 15 of the 23 women (65.2%) with clinical signs, the incidence of a positive test increasing in direct proportion to the amount of yeasts isolated. The test's sensitivity, specificity and predictive values were comparable to those of microscopy and culture. Being both rapid and simple to perform, this new test offers a useful alternative to conventional methods for the diagnosis of vaginal candidosis.     

Evaluation of a latex agglutination kit (Virogen Rotatest) for detection of bovine rotavirus in fecal samples

The performance of the Virogen Rotatest latex agglutination test (LAT) was evaluated for detection of bovine rotavirus antigen. Sixty-three fecal samples from diarrheic calves were collected from November 1999 to May 2000 and screened by LAT, the Rotazyme II enzyme-linked immunosorbent assay (ELISA), and virus isolation (VI) followed by an anti-rotavirus fluorescent-antibody (FA) test to detect the presence of group A rotavirus antigen. Of the 63 samples screened by VI-FA, 33 (58%) tested positive for rotavirus antigen. When the results from the LAT were compared to those from VI-FA, the "gold standard" for detection of bovine rotavirus in fecal samples, the sensitivity and specificity were found to be 87.8 and 73.3%, respectively. Latex agglutination compared with ELISA (the reference method) showed 100% sensitivity and 96.3% specificity, and when ELISA was compared with VI, the sensitivity was 84.8% and the specificity was 73.3%. Latex agglutination is easy to perform in a short time and does not require expensive equipment or skilled personnel, and the reagents have long shelf lives. These factors make the LAT suitable and highly efficient for use in a clinical laboratory as a rapid screening test for bovine rotavirus.