Tulsi D. Chugh

Enterotoxin production by coagulase-negative staphylococci in restaurant workers from Kuwait City may be a potential cause of food poisoning

Staphylococcus aureus and coagulase-negative staphylococci (CNS) were isolated from the hands of food handlers in 50 restaurants in Kuwait City and studied for the production of staphylococcal enterotoxins, toxic shock syndrome toxin-1, slime and resistance to antimicrobial agents. One or a combination of staphylococcal enterotoxins A, B or C were produced by 6% of the isolates, with the majority producing enterotoxin B. Toxic shock syndrome toxin-1 was detected in c. 7% of the isolates; 47% produced slime. In all, 21% of the isolates were resistant to tetracycline and 11.2% were resistant to propamidine isethionate and mercuric chloride. There was no correlation between slime and toxin production or between slime production and antibiotic resistance. The detection of enterotoxigenic CNS on food handlers suggests that such strains may contribute to food poisoning if food is contaminated by them and held in conditions that allow their growth and elaboration of the enterotoxins. It is recommended that enterotoxigenic CNS should not be ignored when investigating suspected cases of staphylococcal food poisoning.

Characterization of rpoB mutations in rifampin-resistant Mycobacterium tuberculosis isolates from the Middle East

The nature and frequency of mutations in the rpoB gene of rifampin-resistant clinical Mycobacterium tuberculosis isolates vary considerably according to geographical locations. There is no information on the prevalence of specific mutations in clinical M. tuberculosis strains isolated from patients in Middle-Eastern countries. In this study, 13 rifampin-resistant and 6 susceptible clinical M. tuberculosis isolates were tested for identification and characterization of mutations in the rpoB gene by INNO-LiPA Rif. TB kit and DNA sequencing of the PCR amplified target DNA. The kit identified all six susceptible strains as rifampin-sensitive and the DNA sequence of the amplified rpoB gene in the target region matched perfectly with the wild-type sequence. The kit identified 12 resistant isolates as rifampin-resistant with specific detection of mutations in 8 isolates while one of the rifampin-resistant strain was identified as rifampin-susceptible. DNA sequencing confirmed these results and, in addition, led to the specific detection of mutations in 4 rifampin-resistant isolates in which specific base changes within the target region could not be determined by the INNO-LiPA Rif. TB kit. The majority (8 of 13) of resistant isolates involved base changes at codon 531 of the rpoB gene. Mutations at codon position 531 within the rpoB gene have also been reported in majority of rifampin-resistant strains from Greece and St. Petersburg, Russia but not from other geographical locations.

Improved detection of mycobacterial antigens in clinical specimens by combined enzyme-linked immunosorbent assays

Three types of antibodies against cellular and secretory-excretory protein antigens were simultaneously used for the direct detection of mycobacterial antigens in sputum and cerebrospinal fluid (CSF) specimens, using enzyme-linked immunosorbent assay (ELISA). The antibodies consisted of in-house raised and prepared anti-whole-cell, heat-killed, and sonicated Mycobacterium tuberculosis, anti-secretory-excretory protein extract of bacilli Calmette-Guerin (BCG) strain, and commercially available anti-BCG. Sputum specimens comprised 24 smear positive, culture positive, and 47 smear-negative, culture positive (SNCP), from patients with pulmonary tuberculosis, as well as 45 smear-negative, culture-negative (SNCN) control samples. The CSF specimens included 18 SNCPs from patients with tuberculous meningitis and 18 SNCN controls. The sensitivity of the individual tests for sputum and CSF specimens ranged from 70% to 79% and 72% to 89%, respectively, whereas in the combined tests it reached 86%-96% for sputum specimens and 100% for CSF specimens. The specificity of ELISAs for sputum specimens was lower in the combined (73%-87%) than in the individual (87%-98%) tests, whereas for CSF specimens it was 100% in all tests. Thus, the combined ELISA approach for mycobacterial antigen detection provides a rapid and reliable laboratory adjunct in the diagnosis of patients with tuberculosis.

Genotypes of Hepatitis C Virus in Kuwait

Hepatitis C virus (HCV) genotypes in 144 patients in Kuwait were determined by the Murex Anti-HCV Serotyping Assay. Twenty-nine (38%) of 77 Kuwaiti patients were found to be infected with genotype 4 and 27% with genotype 1 while of the 41 Egyptian expatriate patients 37 (90%) were infected with genotype 4. Results show that although in native Kuwaiti patients the dominant genotype is 4, other genotypes, especially type 1, occur frequently.

Significance of Atypical Pathogens among Community-Acquired Pneumonia Adult Patients Admitted to Hospital in Kuwait

Objectives: The aim of this study is to determine the microbial etiology and severity of community-acquired pneumonia (CAP) in Kuwait. Subjects and Methods: The severity of consecutive adult CAP cases admitted to 3 hospitals over a 1-year period was classified according to the Pneumonia Outcome Research Team (PORT) severity index. The microbial etiology was determined using standard methods for bacteria and serological tests for atypical and viral pathogens. Results: The study population was 124 of the 135 admissions; 63 female, 61 male; mean age 41.3 ± 18 years. The severity class distribution was: class I 31%, class II 37%, class III 17%, class IV 13%, and class V 2%. Etiological agents were identified from 44 patients (35%), with one pathogen in 31 (25%), two in 9 (7%), and three or more in 4 (3%). The most common pathogens identified were: Mycoplasma pneumoniae in 14 patients (11%), Legionella pneumophila in 10 (8%), Chlamydia pneumoniae in 8 (6%), influenza B virus in 8 (6%), influenza A virus in 5 (4%), Haemophilus influenzae in 4 (3%), Streptococcus pneumoniae in 3 (2%), Staphylococcus aureus in 3 (2%), gram-negative enterobacteria in 5 (4%), Moraxellacatarrhalis in 2 (2%), and viruses in 4 (3%). The yields from laboratory tests were 48% for paired serology, 20% from adequate sputum sample, and 3% from blood culture. Conclusion: Our study shows that a large percentage of mild CAP cases are admitted to hospitals in Kuwait. Atypical pathogens have a significant role in the etiology of CAP. There is overtreatment of CAP with a combination treatment consisting mainly of third-generation chephalosporins and macrolides.

Antimicrobial Susceptibility, Phage Typing and Plasmid Profile of Salmonella enterica Serotype paratyphi A Strains Isolated in Kuwait

Objective: To determine the antimicrobial susceptibility, phage type and plasmid profile pattern of Salmonella enterica serotype paratyphi A strains isolated in Kuwait. Material and Methods: From January 1995 to December 1999, 106 strains of S. enterica serotype paratyphi A isolated from an equal number of cases of enteric fever, attending the Infectious Disease and Mubarak Al-Kabeer Hospitals in Kuwait were investigated. The isolates were tested for antimicrobial susceptibility to 8 commonly used antimicrobial agents. Their phage type and plasmid profile patterns were determined using an international set of phages and Qiagen plasmid mini kit, respectively. Results: All of the isolates were susceptible to ciprofloxacin, cefuroxime, ceftazidime, piperacillin and co-trimoxazole. One hundred isolates were susceptible to ampicillin, 99 to chloramphenicol and 98 to tetracycline. None of the isolates was multidrug resistant. Sixty-six percent of the isolates were phage type I, 27.4% phage type II and 6.6% were untypable. All phage type I and untypable strains had 3 plasmids of 2.2, 5 and 20 kb, whereas phage type II strains had only 1 plasmid of 20 kb. Conclusion: The findings indicate that while all of the isolates of the S. enterica serotype paratyphi A were susceptible to 4 of the drugs tested, some were resistant to ampicillin, chloramphenicol or tetracycline, thereby indicating the need for continued surveillance and monitoring of antimicrobial susceptibility of these isolates.

ELISA in tuberculous meningitis : using two mycobacterial antigens does not improve the diagnostic yield compared to either antigen alone

Abstract The prevention of the devastating sequelae of tuberculous meningitis (TBM) requires the availability of a reliable test for the rapid diagnosis of this condition. This study was carried out by enzyme-linked immunosorbent assay (ELISA) to measure antimycobacterial immunoglobulin (Ig) G, IgM and IgA in the cerebrospinal fluid (CSF) of 11 patients with culture-proven TBM using a whole cell sonicate of heat-killed M. tuberculosis antigen (MTb) and Bacilli-Calmette Guerin (BCG) antigens. The findings were compared to controls consisting of 13 CSF specimens from patients with proven pyogenic meningitis (PM) and 20 control CSF specimens from individuals with normal findings. The ELISA sensitivity, specificity, positive predictive and negative predictive values for IgG were 73%, 91%, 73% and 91%, respectively, against both antigens. The optical density results for IgM and IgA in the CSF of patients with TBM and controls were too low against both antigens to be considered in any analysis. Although ELISA for detection of anti-mycobacterial antibodies in the CSF provides improvement over conventional methods of microscopy and culture, this approach still needs considerable refinement to make it suited for routine clinical use.

Adherence of Staphylococcus epidermidis to Human Pharyngeal Epithelial Cells: Evidence for Lipase-sensitive Adhesin and Glycoprotein Receptor

The adherence of eight strains ofStaphylococcus epidermidis to human pharyngeal epithelial cells (PEC) was investigated. All bacterial strains were nonencapsulated and non-slime producers. Binding was mannose resistant and was not related to surface hydrophobicity and surface charge of bacteria. Adherence to epithelial cells was reduced four- to tenfold (P<0.01) on pretreatment of bacteria with lipase while neuraminidase, phospholipase C, trypsin, and sodium periodate did not alter their binding. The surface carbohydrate profile of bacteria was studied by monitoring adherence to Lectin-Sepharoses. The bacteria did not conform to any pattern, and there was no relation to strain variation. The pretreatment of PEC with trypsin and sodium metaperiodate produced a marked reduction in bacterial binding (three- to 25-fold, P<0.01), but neuraminidase, phospholipase C, and lipase did not have any such effect. These findings provide evidence that the receptors on the surface of PEC are glycoprotein in nature, while the bacterial adhesin is a lipase-sensitive material.

In vitro Activity of Conventional and Newer Antimicrobial Agents against Brucella melitensis Clinical Isolates from Kuwait

The in vitro activity of 15 antimicrobial agents was tested against 21 clinical isolates of Brucella melitensis from Kuwait. The minimum inhibitory concentration for 90% of strains (MIC 90) was determ

Adherence ofStaphylococcus epidermidis to pharyngeal epithelial cells mediated by lipoteichoic acid

Prior treatment of pharyngeal epithelial cells (PEC) with lipoteichoic acid (LTA) derived fromStaphylococcus epidermidis produced a marked inhibition of adherence of the homologous strain and two heterologous strains. The inhibition was dose dependent and saturable with 100 µg/ml of LTA. However, pretreatment of PEC with deacylated LTA did not block the adherence of the three strains tested. A similar but less marked blocking effect on the adherence ofS. epidermidis to PEC was also observed with LTAs derived fromS. aureus andStreptococcus pyogenes. On treatment of bacteria with substances capable of binding to LTA, such as polyclonal mouse anti-LTA antibodies or with human albumin, a marked inhibition of bacterial adherence was observed. Immunofluorescence studies showed that anti-LTA antiserum bound readily to the surface of bacterial cells. These findings provide clear evidence that the lipid component of LTA located on the bacterial surface is centrally involved in the adherence ofS. epidermidis to human mucosal cells.