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M Alfa

Publications and source records attributed to M Alfa.

15 recordsLinked to original sources
PubMed2003

Multilaboratory comparison of proficiencies in susceptibility testing of Helicobacter pylori and correlation between agar dilution and E test methods.

Susceptibility testing was performed at seven Canadian microbiology laboratories and the Helicobacter Reference Laboratory, Halifax, Nova Scotia, Canada, to assess susceptibility testing proficiency and the reproducibility of the results for clarithromycin and metronidazole and to compare the Epsilometer test (E test) method to the agar dilution reference method. Control strain Helicobacter pylori ATCC 43504 (American Type Culture Collection) and 13 clinical isolates (plus duplicates of four of these strains including ATCC 43504) were tested blindly. The National Committee for Clinical Laboratory Standards (NCCLS) guidelines for agar dilution testing were followed, and the same suspension of organisms was used for agar dilution and E test. Antimicrobials and E test strips were provided to the investigators. Methods were provided on a website (www.Helicobactercanada.org). Each center reported MICs within the stated range for strain ATCC 43504. Compared to the average MICs, interlaboratory agreements within 2 log(2) dilutions were 90% (range, 69 to 100%) for clarithromycin by agar dilution, with seven very major errors [VMEs], and 85% (range, 65 to 100%) by E test, with three VMEs. Interlaboratory agreements within 2 log(2) dilutions were 83% (range, 50 to 100%) for metronidazole by agar dilution, with six VMEs and eight major errors (MEs), and 75% (range, 50 to 94%) by E test, with four VMEs and four MEs. At lower and higher concentrations of antibiotic, E test MICs were slightly different from agar dilution MICs, but these differences did not result in errors. When a standardized protocol based on NCCLS guidelines was used, most participants in this study correctly identified clarithromycin- and metronidazole-susceptible and -resistant strains of H. pylori 93% of the time by either the agar dilution or E test method, and the numbers of errors were relatively equivalent by both methods.

Clarithromycin↗
PubMed2001

Multiplex PCR for the detection of tetracycline resistant genes.

Specific primer pairs were selected for the PCR amplification of 14 tetracycline resistant genes commonly found in Gram positive and Gram negative organisms. Combinations of primer pairs were used in multiplex PCR reactions to detect specific groups of tet genes as follows; Group I tet (B), tet (C), tet (D); Group II tet (A), tet (E), tet (G); Group III tet (K), tet (L), tet (M), tet (O), tet (S); Group IV tetA (P), tet (Q), tet (X). To test the multiplex PCR, Groups I and II were used on 25 clinical isolates of Salmonella enterica serovar Typhimurium DT104. Group III primers were used to investigate 19 clinical isolates of methicillin-resistant Staphylococcus aureus. Multiplex PCR should result in significant savings in terms of labour and cost in analysis of a large number of strains when compared with using an individual PCR for targeting each gene. It may also be a useful method to differentiate the types of tetracycline resistance when used as an additional marker for the purpose of outbreak investigation and surveillance.

DNA Fingerprinting↗
PubMed2001

Development of a Canadian standardized protocol for subtyping methicillin-resistant Staphylococcus aureus using pulsed-field gel electrophoresis.

A panel of 24 methicillin-resistant Staphylococcus aureus strains was distributed to 15 laboratories in Canada to evaluate their in-house pulsed-field gel electrophoresis (PFGE) protocols and interpretation criteria. Attempts to compare fingerprint images using computer-aided analysis were not successful due to variability in individual laboratory PFGE protocols. In addition, individual site interpretation of the fingerprint patterns was inadequate, as 7 of 13 sites (54%) made at least one error in interpreting the fingerprints from the panel. A 2-day standardized PFGE protocol (culture to gel image) was developed and distributed to all of the sites. Each site was requested to use the standardized protocol on five strains from the original panel. Thirteen sites submitted gel images for comparisons. The protocol demonstrated excellent reproducibility and allowed interlaboratory comparisons with Molecular Analyst DST software (Bio-Rad) and 1.5% band tolerance.

Bacterial Typing Techniques↗
PubMed2000

Analysis of microbiological trends in peritoneal dialysis-related peritonitis from 1991 to 1998.

The microbial cause of peritoneal dialysis-related peritonitis is an important determinant of clinical outcome and the basis of widely used treatment guidelines. Five hundred forty-six cases of peritonitis in 374 patients from 1991 to 1998 were analyzed. The rate of peritonitis declined significantly from 1.37 episodes/patient-year in 1991 to 0.55 episode/patient-year in 1998 (P = 0.02). The rate of Gram-positive peritonitis decreased significantly from 0.75 to 0.28 episode/patient-year during the same period (P = 0.02). Conversely, the occurrence of Gram-negative peritonitis remained constant at approximately 0.16 episode/patient-year (P = 0.28). Staphylococcus epidermidis and Staphylococcus aureus were the most common causes of peritonitis, isolated in 27.8% and 19.3% of the culture-positive cases, respectively. A distinct decrease in peritonitis caused by S epidermidis was observed, with 0.40 episode/patient-year in 1991 compared with 0.11 to 0.20 episode/patient-year during subsequent years. The rate of infections caused by S aureus decreased significantly over time from a high of 0.21 episode/patient-year in 1992 to a low of 0.04 episode/patient-year in 1998 (P = 0.01). Pseudomonas aeruginosa, Escherichia coli, and KLEBSIELLA: species were the most common causes of Gram-negative peritonitis, identified in 7.1%, 6.8%, and 5.2% of culture-positive cases, respectively. The most dramatic increase in antibiotic resistance was seen among S epidermidis. From 1991 and 1992 to 1997 and 1998, resistance to ciprofloxacin increased from 5.4% to 47.8% (P = 0.003), and resistance to methicillin increased from 18.9% to 73.9% (P = 0.03). Our study showed significant trends in the causative pathogens of peritoneal dialysis-related peritonitis and dramatic increases in antibiotic resistance. These data support further study and warrant reevaluation of current treatment practices.

Case-Control Studies↗
PubMed2000

Mycobacterium branderi from both a hand infection and a case of pulmonary disease.

Mycobacterium branderi, a potential human pathogen first characterized in 1995, has been isolated from respiratory tract specimens. We report here a case in which M. branderi was the only organism isolated upon culture from a hand infection. This isolate, along with a second isolate from a bronchial specimen, was subjected to conventional identification tests for mycobacterial species. Further analysis by high-performance liquid chromatography (HPLC) of mycolic acids and 16S rRNA gene sequencing was performed, and the antibiotic susceptibility profile was determined for both strains. Biochemical tests and the HPLC pattern were consistent with that of M. branderi and M. celatum, which are very similar. The 16S rRNA gene sequence of both strains corresponded to that of M. branderi and enabled us to confidently differentiate this organism from other closely related species such as M. celatum. This contributes to a further understanding of the status of this species as a potential human pathogen as well as illustrating the need for molecular diagnostics as a complementary method for the identification of rare mycobacterial species.

Chromatography, High Pressure Liquid↗
PubMed1996

A morphological study of penile chancroid lesions in human immunodeficiency virus (HIV)-positive and -negative African men with a hypothesis concerning the role of chancroid in HIV transmission.

Chancroid, the most common cause of genital ulceration in Africa, is known to be associated epidemiologically with heterosexual transmission of human immunodeficiency virus (HIV). The pathophysiological mechanisms by which chancroid might facilitate the spread of HIV are obscure. To investigate the role of chancroid in HIV transmission, the authors studied the histological features of biopsies from 11 men with penile chancroid lesions including five who were serologically positive for HIV. The histomorphologic and immunophenotypic nature of the inflammatory infiltrates suggests that there is a significant role for cell-mediated immunity in the host response to Hemophilus ducreyi infection. This response may be critical to the role of chancroid in HIV transmission.

Black or African American↗
PubMed1995

Continuous quality improvement for introduction of automated blood culture instrument.

Despite the critical nature and high cost of blood cultures, hospitals rely on manufacturers' test site data. As a result, in-hospital testing and compliance evaluation of newly acquired instruments are seldom done. The goal of this study was to apply a continuous quality improvement approach and to develop assessment criteria for all stages from the purchase order, through the on-site instrument evaluation, to the compliance evaluation. Despite the introduction of an automated high-blood-volume instrument (BacT/Alert) in our hospital, 56% of adult patients had only one venipuncture and 89.5% had < or = 20 ml of total blood volume sampled. False positives were associated with overfilling of bottles. These problems occurred because the phlebotomists did not like to perform multiple venipunctures on ill patients; therefore, they were drawing 20 ml of blood from one venipuncture and splitting it between two bottles. Unknown to the staff, the vacuum in the bottles draws significantly more than 10 ml of blood; therefore, the first bottle in the set was frequently overfilled and the second bottle was frequently underfilled. A diagrammatic guideline for a new blood culture protocol based on two venipunctures, taken one immediately after the other, to inoculate three bottles was developed. Compliance evaluation demonstrated that within 1 month of starting the new protocol, 74% of patients had at least two or more venipunctures and 60% had > or = 30 ml of blood drawn per patient episode. This study demonstrates the need for continuous quality improvement, including compliance evaluation, to ensure that the potential benefits of newer blood culture technology are actually realized.

Adult↗
PubMed1995

Fast food arthritis--a clinico-pathologic study of post-Salmonella reactive arthritis.

OBJECTIVE: To study the clinical presentation, immunogenetics, and serum immune response to lipopolysaccharide (LPS) in a cohort of patients with post-Salmonella reactive arthritis (ReA). METHODS: A validate ReA screening questionnaire (Quest 2) was mailed to 919 individuals reporting symptoms of gastroenteritis to the health department after eating at a single restaurant. Three hundred twenty-one persons returned questionnaires; 170 reported symptoms outside the gastrointestinal tract; 23 of those 170 reporting persistent joint symptoms were seen 4 to 16 weeks after the outbreak and 5 of the 23 were seen in followup 12 to 20 weeks later. Clinical features, HLA Class I typing, serum soluble CD8 levels, and serum antibodies to gram negative LPS by ELISA were determined. RESULTS: Joint complaints were reported more frequently by individuals with a longer duration of diarrhea. Upper extremity joints were frequently involved, and 66% reported one or more extraarticular symptoms of Reiter's syndrome. Three of 5 typed individuals were HLA-B27 positive, including 3 of the 4 most severely involved. Serum soluble CD8 levels correlated poorly with disease activity measured either clinically or by C-reactive protein. Antibodies to Klebsiella and Shigella LPS rose over time, while antibodies to Salmonella LPS fell. CONCLUSION: The clinical picture of post-Salmonella ReA is less stereotyped than often assumed, although severity correlated with HLA-B27 status. The association of joint symptoms with duration of diarrhea and the kinetics of the anti-LPS antibody response support the hypothesis that abnormal gut permeability plays a role in the pathogenesis of post-Salmonella ReA.

Antibodies, Bacterial↗
PubMed1994

Utilization of commercial antisera and flow cytometry in HLA-B27 typing.

The study of large groups of individuals (from reactive arthritis epidemics, for example) for epidemiologic and genetic data has been hampered by the techniques by which HLA typing is performed. Microlymphocytotoxicity is costly not only in materials but also in the time required to perform a given assay and thus does not lend itself to large scale testing. To study such epidemics, in the best way, one must be able to respond as soon as possible after the epidemic is recognized and on a large scale. We thus examined HLA-B27 typing by flow cytometry and commercially available monoclonal antibodies. Two commercially available monoclonal HLA-B27 antibodies were compared. We conclude that the utilization of monospecific HLA-B27 antibodies and flow cytometry allows for the determination of HLA-B27 and HLA-B7 phenotype of a large number of individuals over a short period of time, thereby making the study of a large group both economical and feasible.

Arthritis, Reactive↗
PubMed1994

Methicillin-resistant Staphylococcus aureus in tertiary care institutions on the Canadian prairies 1990-1992.

OBJECTIVE: To review experience with methicillin-resistant Staphylococcus aureus (MRSA) in tertiary acute-care teaching hospitals on the Canadian prairies. DESIGN: Retrospective review for a 36-month period, 1990 through 1992. SETTING: Five tertiary acute-care teaching hospitals in three Canadian prairie provinces. METHODS: MRSA isolates and susceptibility were identified through the clinical microbiology laboratory at each institution. For each patient, data collected included duration of institutional residence prior to isolation, patient ethnic background, age, sex, and antimicrobial susceptibility. Epidemiologic typing of strains used restriction fragment length polymorphism analysis by pulsed-field gel electrophoresis. RESULTS: Two hundred fifty-nine MRSA isolates were identified in 135 patients during the 36 months, with substantial institutional variation in number of isolates. No consistent increase in yearly numbers of isolates was apparent. Patients usually had MRSA identified at admission (62%); only one of five centers had the majority of isolates acquired nosocomially. Patients with MRSA present at admission were more frequently of aboriginal (First Nations) ethnicity (62% compared with 14% of nosocomial; P < 0.001). Pulsed-field gel electrophoresis of 167 isolates from 135 patients revealed 46 different strains with little interprovincial or interinstitutional identity of strains. CONCLUSIONS: MRSA isolated in patients in tertiary care institutions in these three Canadian provinces usually is acquired prior to admission. A disproportionate number of isolates are identified in aboriginal Canadians. Epidemiologic typing was consistent with a polyclonal origin of MRSA in this geographic area.

Adult↗
PubMed1994

Serologic testing for reactive arthritis.

The objective of the study was to determine the sensitivity and specificity of quantitative serum antibody response to Salmonella enteritidis lipopolysaccharide (LPS) as a diagnostic test for post-Salmonella reactive arthritis (ReA). In a single food-source outbreak of Salmonella enteritidis, serum was collected from dysenteric individuals with and without ReA at 6, 12, and 24 months post infection. Serum was also collected from control patients with no prior exposure to Salmonella infection. Quantitative measurements of isotypic antibodies to Salmonella enteritidis LPS were performed by an ELISA. Sensitivity and specificity of quantitative isotypic antibody levels over time were plotted on receiver operator characteristic (ROC) curves. Serum IgG and IgA anti-LPS were found to be present in higher levels in the ReA patients than in controls. Using the optimal cutoff of 0.10 selected from an ROC curve, IgG anti-LPS is 88% sensitive and 94% specific, and IgA anti-LPS is 75% sensitive and 100% specific. We conclude that IgA anti-LPS is both sensitive and specific in distinguishing prior exposure to Salmonella LPS in ReA patients compared to unexposed controls.

Antibodies, Bacterial↗
PubMed1994

Secretory immune response and clinical sequelae of Salmonella infection in a point source cohort.

OBJECTIVE: To determine the kinetic isotypic serum and secretory immune response to Salmonella enteritidis in a cohort of individuals exposed to the organism in a single food source outbreak of dysentery. To determine the clinical outcome and immunogenetics of the exposed cohort and to correlate these features with the immune response. METHODS: Following a single point source outbreak of Salmonella enteritidis, a cohort of dysenteric individuals were ascertained using a reactive arthritis screening questionnaire (QUEST). Serum and stimulated saliva samples were obtained at 6, 12, and 24 months following the outbreak of dysentery; examinations were conducted at the same time. Two unexposed control groups were ascertained: (1) general rheumatology clinic patients and (2) well nonarthritic family practice patients. An ELISA to determine quantitative IgA responses to Salmonella enteritidis lipopolysaccharide (LPS) was performed. RESULTS: Eleven of the 84 exposed individuals with dysentery developed reactive arthritis (ReA) of reactive enthesitis (ReE). There was a prolonged salivary IgA anti-LPS response in both the ReA/ReE and DYS (dysentery alone) patients compared with unexposed controls. A ratio of salivary IgA anti-LPS/serum IgA anti-LPS > 1 was associated with a good outcome (remission) of ReA, whereas a ratio < 1 was associated with chronic disease. CONCLUSIONS: There is a more prolonged humoral immune response to Salmonella LPS in exposed individuals than hitherto described. A risk factor in the prolongation of ReA is the inability to mount an appropriate specific salivary (secretory) immune response.

Adolescent↗
PubMed1982

A transport method for swab specimens submitted for aerobic and anaerobic bacteriology.

The need for separate swab transport methods for aerobes and anaerobes may result in inadequate transport of specimens for anaerobic bacteriology. Most microbiology laboratories in Australia rely on Stuart's transport medium to protect anaerobic bacteria. This paper presents a new, simple transport medium (Transport Deep) suitable for sue with aerobes and anaerobes. Comparative evaluations demonstrate that Transport Deep is as good as Stuart's medium for the maintenance of fastidious bacteria and is far superior for the protection of even extremely oxygen-sensitive anaerobes. This medium has been used successfully in a large Sydney hospital for more than a year. It is proposed that Transport Deep be used on a routine basis for all swab specimens.

Aerobiosis↗
PubMedDate not supplied

Morphology of the cells and colonies of Mycoplasma hominis.

Studies of the morphology Mycoplasma hominis have shown that the cells resemble those of small coccoid or ovoid bacteria; the microstructure of the organism is compatible with reproduction by a fission process. Although dividing forms with equal-sized lobes are common, fragmenting filaments and replicating buds sometimes occur, perhaps in response to suboptimal growth conditions. Data based on morphometric analysis show that cells of M. hominis ATCC 14027 have average dimensions of 0.27 X 0.74 micron and a mean diameter of 0.42 micron. The volume of a M. hominis cell was only about 60% of that of Ureaplasma urealyticum T960, a species that has a mean cell diameter of 0.5 micron and that is sometimes a pathogen of the human genital tract. There are wide variations in the colonial morphology of M. hominis.

Bacteriological Techniques↗