MATERIALS AND METHODS- Urinary tract infection among HIV
MATERIALS AND METHODS
From June 2006 to November 2007, 342 mid-stream clean catched urine samples were collected and processed at the bacteriology laboratory of Benghazi Center of Infectious Diseases and Immunity. Samples were obtained from female and male adults (>12 years of age) and children (< 12 years of age) represented by 137 inpatients and 205 outpatients with HIV infection. Urine samples were immediately streaked with a calibrated loop onto Cysteine Lactose Electrolytes DeficiencyMedium(CLED) plates (Oxoid, Ltd., Basingstoke, Hampshire, England U.K.) then incubated at 35-37°C aerobically overnight.
For urinalysis sediment microscopy, 10 ml amounts of urine specimens were centrifuged at 2000 rpm for 5 minutes. White blood cells and bacteria per high-powered filedwerecountedand compared with urine culture results. The urine dipstick testing, chemstrip 9 test strips (Medi-Test Combi 9 Macherey-Nagel, German) were used according to the manufacture`s instruction.
Identificationofbacterialisolateswascarried out using standardized microbiological techniques including: colony morphology, gram stain, API 20E system (bioMeriuex, France), Catalase test, Staphylase test (Oxoid, Ltd, UK), Streptococcal grouping kit (Oxoid Ltd, UK), DNase medium (Oxoid Ltd, Basingstoke, Hampshire, England), Mannitol salt agar (Oxoid Ltd, Basingstoke, Hampshire, England), Novobiocin (Abtek, Biologicals Ltd, Liver pool, UK). The disc diffusion technique (3) with Turbidity standard of 0.5 McFarland was used for antibiotic sensitivity testing using Muller-Hinton agar (Oxoid Ltd, Basingstoke, Hampshire, England). Isolates were tested against the following antibiotics: Ampicillin (10μg), Amoxycillin/clavulanic acid (30μg), ciprofloxacin(5μg),Chloramphenicol(30μg), Nitrofurantoin (300μg), Imipenem (10μg), Gentamicin (10μg), Amikacin (30μg), Kanamicin (30μg), Trimethoprim/sulfamethoxazole (25μg), Tetracycline (30μg), Nalidixic acid (30μg). all antimicrobial tested were from Oxoid` UK.
DEFINITIONS:
A positive urine culture was definedaspuregrowth of a urinary tract pathogen exceeding
100,000 CFU/mL. Cultures with growth of 500 CFU/mL of mixed organisms or non-pathogens were considered contaminated. No growth (<100 CFU/mL) or growth of <500 CFU/mL was considered negative. Significantpyuriawas definedbythepresenceof>5whitebloodcells per high-power field(4,5).Bacteriuriawas definedasmanybacteriaseenunderHPF.Intermediate and resistant strains were grouped together and classifiedasresistant.
RESULTS
Out of the 342 mid stream clean catch urine specimens tested; there were 25 cultures positive (7.3%). Overall, there were more females infected with UTI than males with 18 (72%) of cases among females. The distribution of UTI cases among the study group is described in Table 1. Klebsiella species was the most frequently isolated pathogen. Nine (36%) isolates were identifiedas6K. pneumoniae and 3 K. terrigena followed by 8 (32%) E. coli; 3 (12%) staphylococcus aureus; 2 (8%) Staphylococcus epidermidis; and 1 (4%) each of Staphylococcus saprophyticus; Enterobacter aerogenes; Citrobacter koseri and Proteus mirabilis.
Among 25 positive cultures, only one specimen was positive for nitrite test, pyuria in 15 specimens, and bacteriuria in 3 specimens (Table 2). Significantpyuriawaspresentin8specimens (25.8%) of contaminated cultures and in 25 specimens (8.7 %) of negative cultures as seen in Tables 2 and 3. Bacteriuria in 6 specimens (19.4 %) of contaminated cultures
From June 2006 to November 2007, 342 mid-stream clean catched urine samples were collected and processed at the bacteriology laboratory of Benghazi Center of Infectious Diseases and Immunity. Samples were obtained from female and male adults (>12 years of age) and children (< 12 years of age) represented by 137 inpatients and 205 outpatients with HIV infection. Urine samples were immediately streaked with a calibrated loop onto Cysteine Lactose Electrolytes DeficiencyMedium(CLED) plates (Oxoid, Ltd., Basingstoke, Hampshire, England U.K.) then incubated at 35-37°C aerobically overnight.
For urinalysis sediment microscopy, 10 ml amounts of urine specimens were centrifuged at 2000 rpm for 5 minutes. White blood cells and bacteria per high-powered filedwerecountedand compared with urine culture results. The urine dipstick testing, chemstrip 9 test strips (Medi-Test Combi 9 Macherey-Nagel, German) were used according to the manufacture`s instruction.
Identificationofbacterialisolateswascarried out using standardized microbiological techniques including: colony morphology, gram stain, API 20E system (bioMeriuex, France), Catalase test, Staphylase test (Oxoid, Ltd, UK), Streptococcal grouping kit (Oxoid Ltd, UK), DNase medium (Oxoid Ltd, Basingstoke, Hampshire, England), Mannitol salt agar (Oxoid Ltd, Basingstoke, Hampshire, England), Novobiocin (Abtek, Biologicals Ltd, Liver pool, UK). The disc diffusion technique (3) with Turbidity standard of 0.5 McFarland was used for antibiotic sensitivity testing using Muller-Hinton agar (Oxoid Ltd, Basingstoke, Hampshire, England). Isolates were tested against the following antibiotics: Ampicillin (10μg), Amoxycillin/clavulanic acid (30μg), ciprofloxacin(5μg),Chloramphenicol(30μg), Nitrofurantoin (300μg), Imipenem (10μg), Gentamicin (10μg), Amikacin (30μg), Kanamicin (30μg), Trimethoprim/sulfamethoxazole (25μg), Tetracycline (30μg), Nalidixic acid (30μg). all antimicrobial tested were from Oxoid` UK.
DEFINITIONS:
A positive urine culture was definedaspuregrowth of a urinary tract pathogen exceeding
100,000 CFU/mL. Cultures with growth of 500 CFU/mL of mixed organisms or non-pathogens were considered contaminated. No growth (<100 CFU/mL) or growth of <500 CFU/mL was considered negative. Significantpyuriawas definedbythepresenceof>5whitebloodcells per high-power field(4,5).Bacteriuriawas definedasmanybacteriaseenunderHPF.Intermediate and resistant strains were grouped together and classifiedasresistant.
RESULTS
Out of the 342 mid stream clean catch urine specimens tested; there were 25 cultures positive (7.3%). Overall, there were more females infected with UTI than males with 18 (72%) of cases among females. The distribution of UTI cases among the study group is described in Table 1. Klebsiella species was the most frequently isolated pathogen. Nine (36%) isolates were identifiedas6K. pneumoniae and 3 K. terrigena followed by 8 (32%) E. coli; 3 (12%) staphylococcus aureus; 2 (8%) Staphylococcus epidermidis; and 1 (4%) each of Staphylococcus saprophyticus; Enterobacter aerogenes; Citrobacter koseri and Proteus mirabilis.
Among 25 positive cultures, only one specimen was positive for nitrite test, pyuria in 15 specimens, and bacteriuria in 3 specimens (Table 2). Significantpyuriawaspresentin8specimens (25.8%) of contaminated cultures and in 25 specimens (8.7 %) of negative cultures as seen in Tables 2 and 3. Bacteriuria in 6 specimens (19.4 %) of contaminated cultures
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