Urinary tract infection (UTI) results from the presence and multiplication of microorganisms in one or more structures of the urinary tract. Protection against infection is normally given by the constant flow of urine and regular bladder emptying. Infection may spread to surrounding tissues (e.g. Perinephric abscess) or to the bloodstream.
Urinary tract infection (UTI) results from the presence and multiplication of microorganisms in one or more structures of the urinary tract. Protection against infection is normally given by the constant flow of urine and regular bladder emptying. Infection may spread to surrounding tissues (e.g. Perinephric abscess) or to the bloodstream.
Urinary tract infection (UTI) results from the presence and multiplication of microorganisms in one or more structures of the urinary tract. Protection against infection is normally given by the constant flow of urine and regular bladder emptying. Infection may spread to surrounding tissues (e.g. Perinephric abscess) or to the bloodstream.
1. BACKGROUND .................................................................................. 3 2. PURPOSE ........................................................................................... 4 3. RESPONSIBILITIES ........................................................................... 4 4. PROCEDURE ..................................................................................... 4 4.1 MATERIALS NEEDED ................................................................... 4 4.2 STORAGE CONDITIONS .............................................................. 4 4.3 TEST PROCEDURE ...................................................................... 4 4.4 QUALITY CONTROL ...................................................................... 5 4.5 INTERPRETATION OF RESULTS ................................................. 5 4.6 LIMITATIONS ................................................................................. 6 5. HEALTH & SAFETY ........................................................................... 6 6. ASSOCIATED PROCEDURES .......................................................... 6 7. REFERENCES .................................................................................... 7 8. APPENDICES ..................................................................................... 8 Appendix 1: Fastread 102 counting chamber instructions for use .... 8 9. Training Record for this SOP ......................................................... 10
MLW.SOP.L.MICRO.016 Version 1 Page 3 of 10 1. BACKGROUND Urinary tract infection (UTI) results from the presence and multiplication of microorganisms in one or more structures of the urinary tract with associated tissue invasion. This can give rise to a wide variety of clinical syndromes. These include acute and chronic pyelonephritis (kidney and renal pelvis), cystitis (bladder), urethritis (urethra), epididymitis (epididymis) and prostatitis (prostate gland). Infection may spread to surrounding tissues (e.g. perinephric abscess) or to the bloodstream.
Protection against infection is normally given by the constant flow of urine and regular bladder emptying. Urine is a poor culture medium for many bacteria due to its acidity, high urea concentration and variable osmolality and, in men, possibly partly as a result of antibacterial activity of prostatic secretions 1 .
Bacteriuria Implies that bacteria are present and may be cultured from urine. The patient may or may not be symptomatic. Symptomatic patients They may be bacteriuric or abacteriuric. Symptoms in children and the elderly, when present, may be non-specific and difficult to interpret. Frequency The average bladder capacity is about 500 mL. Significant reduction in capacity accompanies acute inflammation which can lead to an increase in the frequency of micturition. Dysuria Painful and difficult micturition. Urgency A strong desire to empty the bladder, which can lead to incontinence. Nocturia Waking in the night one or more times to void the bladder 2 . Nocturnal enuresis The involuntary voiding of urine during sleep, i.e. bed-wetting. Incontinence The involuntary leakage of urine. The commonest form of this is stress incontinence where leakage accompanies an increase in intra-abdominal pressure due to sneezing, coughing or laughing. Overflow or dribbling incontinence accompanies an overfilled bladder. Prostatism Symptoms include: hesitancy or delay in initiating micturition, intermittency or interruption, and reduced force of the urine stream resulting from prostate gland pathology. Renal colic This is characterised by very severe cramping pain resulting from distension of the ureter and pelvis above an obstruction such as a renal stone. Often accompanied by frequency and urgency.
MLW.SOP.L.MICRO.016 Version 1 Page 4 of 10 Asymptomatic bacteriuria Common in several patient groups, particularly the elderly, pregnant women and diabetic patients.
2. PURPOSE This SOP explains how to perform Urine microscopy and culture on mid-stream urine samples received at the MLW laboratory. If any other type of urine specimen is received this should only be processed after discussion with, and advice from, the clinical microbiologist.
4.2 STORAGE CONDITIONS 1) Agar plates should be stored at 2-8oC and used within 1 month of their production. 2) All other materials are stored at room temperature
4.3 TEST PROCEDURE
Day one: 1) Gently mix the urine sample by inverting the tube several times (do not shake the sample)
MLW.SOP.L.MICRO.016 Version 1 Page 5 of 10 2) Perform a cell count using a Fastread 102 counting chamber (see appendix 1) 3) Place 1 drop of urine using a pipette into the sample application area of the chosen counting well and allow it to distribute evenly throughout the counting area. 4) Count the numbers of cells (white cells and red cells) across all ten large grids (4x4 small grids) to achieve a count per l or count a smaller area and do the appropriate calculation to achieve a count per l. 5) To achieve a count per ml multiply the count per l by 1000. 6) Inoculate 1l of urine sample onto a blood agar plate and a CLED agar plate using a 1l sterile loop and spread to achieve single colony growth 7) Incubate both plates in air at 37 o C overnight
Day two: 1) Check both plates for bacterial growth and identify the organisms according to interpretation of results (4.5) below. 2) Perform sensitivity tests on significant bacterial growth (as defined by the table in interpretation of results below) using MLW.SOP.MICRO.208.
Day three: 1) Read and interpret sensitivity tests as describe in MLW.SOP.MICRO.208
4.4 QUALITY CONTROL See associated procedures
4.5 INTERPRETATION OF RESULTS Identification of bacterial growth and subsequent sensitivity testing should only be done if the bacterial growth is considered significant in conjunction with the microscopic analysis (see table below).
White cell count Culture result (per agar plate) Significant
Any Pure single organism 10 colonies Yes
MLW.SOP.L.MICRO.016 Version 1 Page 6 of 10 Any Pure single organism <10 colonies No 10 5 per ml 2 types of organisms 10 colonies Yes 10 5 per ml More than 2 types of organisms 10 colonies No <10 5 per ml Mixed culture 10 colonies No Any Mixed culture < 10 colonies No
Reporting: Bacterial Growth Report as:
GNB, Yellow on CLED Lactose fermenting coliform GNB, Clear on CLED Identify (MLW.SOP.MICRO.204) GPC Identify (MLW.SOP.MICRO.203) Mixed culture (not significant) No significant growth No growth No bacterial growth
4.6 LIMITATIONS 1) Mid-stream urines give more reliable microscopy results 2) The urine must be tested within 24 hours of collection as contaminating bacterial overgrowth may obscure significant organisms after this time unless collected into a sample tube containing boric acid preservative. 3) Urine samples recovered from nappies are NOT suitable for examination
5. HEALTH & SAFETY 1) Follow health and safety guidelines as laid down in the MLW Laboratory Health and Safety Manual