Urinalysis Standard Reporting Result Using Urine Flow cytometry

Dr. Eirene Jaquelene K.T., SpPK

OUTLINE

Introduction

Principle of UF-4000

Evaluation study of urine sediment reference range using UF-4000

INTRODUCTION

• Is a labor-intensive, time consuming test• Lack of standardization in high-volume laboratories• Variations in the test results due to skills of technician

Manual urine sediment test

• Made urine sediments tests more efficient, less laborand more objective allows a better standardization of urine sediment analysis

Automated urine flow cytometry

• Is an automated flow cytometer for urine sediments analysis new improvement i.e blue semiconductor laser and depolarized side scatter light.

UF-4000

INTRODUCTION

• provides quantitative results of urine sediments either in microliter (μL), high performance field (HPF) and low performance field (LPF). This instrument doesn’t have reference values for Indonesian population.

UF-4000

• Are important for interpretation of laboratory results. Reporting it must be technically correct and must provide unambiguous information.

Reference values

•Recommends that every laboratory established a reference values for its own population.CLSI

PRINCIPLE OF UF-4000

Flow cytometry

Blue laser & depolarized side scatter

light

Waveform analysis

SFch & CRch

Taken from Sysmex 2013

PRINCIPLE OF UF-4000

Taken from Sysmex 2013

PRINCIPLE OF UF-4000 Taken from Sysmex 2013

PRINCIPLE OF UF-4000 Taken from Sysmex 2013

EVALUATION STUDY OF URINE SEDIMENT USING UF-4000

AIM OF STUDY:• To evaluate reference range of urine sediments analysis

parameters (RBC, WBC, Epithelial Cells, Types of Epithelial Cells and Bacteria) using UF-4000

• To evaluate reference range of research parameters (Cast, Hyaline cast, Path. Cast, Crystal, Yeast, Sperm and Mucus) using UF-4000

METHOD

Time of study : October 2018 – April 2019

Place of study :

RS Bhayangkara tingkat I Raden Said Sukanto

Study population: participants of medical check up who fulfilled inclusion criteria

Amount of samples needed :

o To establish reference range : 120 female, 120 male

METHOD Inclusion criteria:

Random urine samples from participants of medical check up which considered clinically healthy.

Female and male, age 18 – 65 year old

Fresh urine collected with clean catch mid stream

Normal urinalysis, CBC, random blood glucose, liver (AST,ALT) and renal (urea, creatinine) function results

Exclusion criteria:

Female on menstrual period and pregnant

Abnormal in either urinalysis CBC, RBG, Liver function (AST,ALT), renal function (urea, creatinine) results

Data analysis : collected data were analysed using Medcalc

OPERATIONAL LIMITS

Random fresh urine : Clean-catch midstream urine samples that are examined within 2 hours after collection.

Clinically healthy : medical check up participants with no health complaints during check up, urinalysis, complete blood count (CBC) , random blood glucose, liver function (AST,ALT), renal function (urea, creatinine) results are within reference range

OPERATIONAL LIMITS

Normal urinalysis:

Macroscopic : yellow/straw, clear

Urine chemistry : pH 5.0 – 8.0, SG 1.003 – 1.030, Glucose (-), Bilirubine (-), blood (-), protein (-), urobilinogen < 2 mg/dl or 34 μmol/L, leukocyte esterase (-)

EVALUATION PROCEDUREParticipants of medical check up

Participants were educated to collect mid stream urine samples and blood were drawn by phlebotomists

Random urine were collected in urine container

Urine were analysed using Sysmex UN-2000 (UC-3500 and UF-4000), Blood were analysed for CBC, RBG, liver function (AST,ALT), renal function (urea, creatinine)

Normal urinalysis, CBC, RBG, renal and liver function. Age of participants 18 – 65 yo

Abnormal results of urinalysis/CBC/RBG/renal/liver function, participants age are not within 18 – 65 yo

Data from UF-4000 were collected Excluded

Analysed with Medcalc

Parameter Unit Female (n=120) Male (n=120)Mann-Whitney

pAnalysis Reference range Reference range

Red blood cells/uL 0.20 - 10.10 0.10 - 6.60 <0.0001

/HPF 0.00 - 1.80 0.00 - 1.10 0.0002

White blood cells/uL 0.10 - 6.55 0.10 - 6.55 0.8131

/HPF 0.00 - 1.10 0.00 - 1.10 0.8113

Epithelial cells/uL 0.10 - 7.81 0.10 - 2.94 <0.0001

/HPF 0.00 - 1.42 0.02 - 0.53 <0.0001

Squamous EC/uL 0.00 - 5.80 0.00 - 1.40 <0.0001

/HPF 0.00 - 1.00 0.00 - 0.20 <0.0001

Parameter Unit

Female MaleMann-Whitney

pAnalysis Reference range Reference range

Transitional EC/uL

0.00 - 0.20 0.00 - 0.20 0.1504

/HPF0.00 - 0.10 0.00 - 0.00 0.0067

RTEC/uL

0.00 - 2.05 0.00 - 2.05 0.843

/HPF0.00 - 0.50 0.00 - 0.30 0.0055

Bacteria /uL1.05 - 107.82 0.06 - 58.23 <0.0001

/HPF0.19 - 19.42 0.01 - 10.48 <0.0001

ParameterUnit

Female MaleMann-Whitney

pResearch Reference range Reference range

Cast/uL 0.00 - 0.40 0.00 - 0.40 0.5334

/LPF 0.00 - 1.20 0.00 - 1.20 0.3962

Hyaline cast/uL 0.00 - 0.41 0.00 - 0.41 0.2219

/LPF 0.00 – 1.20 0.00 - 1.20 0.1625

Pathologic cast

/uL 0.00 - 0.13 0.00 - 0.00 0.0037

/LPF 0.00 - 0.40 0.00 - 0.00 0.0037

ParameterUnit

Female MaleMann-Whitney

pResearch Reference range Reference range

Crystal/uL 0.00 - 0.00 0.00 - 0.00 1

/HPF 0.00 - 0.00 0.00 - 0.00 1

Yeast/uL 0.00 - 0.70 0.00 - 0.20 0.001

/HPF 0.00 - 0.10 0.00 - 0.00 <0.0001

Sperm/uL 0.00 - 0.00 0.00 - 0.00 1

/HPF 0.00 - 0.00 0.00 - 0.00 1

Mucus/uL 0.00 - 0.78 0.00 - 0.78 0.0681

/LPF 0.00 - 1.20 0.00 - 2.80 0.0433

DISCUSSIONS

RBC count in female > male, the upper value in female is 10.10/μL (1.80/HPF); male 6.60/ μL (1.10/HPF). These values are similar to reference values from existing literature which are 1-2/HPF and <3/HPF.

The upper values of WBC in male & female in this study is 6.55/μL (1.10/HPF). This result is within WBC count in normal urine from other literature which are <2/HPF and <5/HPF.

DISCUSSIONS

Epithelial cells (EC) results in female > male, the upper limit are 7.81/μL (1.42/HPF) in female and 2.94/μL (0.53/HPF) in male.

The epithelial cells in the urine may originate from any site in the genitourinary tract. Normally, a few cells from these sites can be found in the urine as a result of the normal sloughing off of old epithelial cells.

DISCUSSIONS Three types of epithelial cells are seen in urine: squamous,

transitional (urothelial), and renal tubular.

Squamous EC in female > male. The upper limits are 5.80/ μL (1/HPF); male 1.40/μL (0.2/HPF). Usually at least few squamous EC are normally present in urine sediment.

Transitional EC originate from the lining of ureters, and bladder. In this study showed the upper limit in male is 0.20/μL (0.0/HPF) and 0.20/μL (0.1/HPF) in female.

DISCUSSIONS RTEC are very rare/few in normal urine. The presence of > 2RTE/HPF

or >15/10 HPF indicates tubular injury. This study found that upper limits in female is 2.05/μL (0.5/HPF) and 2.05/μL (0.3/HPF) in male.

Bacteria results showed higher bacteria in female. The upper limit are 107.82/μL (19.42/HPF) for female and 58.23/μL (10.48/HPF) in male.

Smith et al. found bacteriuria > 105 CFU/ml or 100/μL is accepted as indicative of infection. Manoni et al. found bacteriuria > 125/μL as a cut off for screening of UTI.

DISCUSSIONS

Terajima et al. explained that higher bacteria in female urine could be caused by contamination of lactobacilli from vaginal secretion.

Cast results are the same in female & male i.e 0.40/μL (1.2/LPF). Hyaline cast results are also the same in both gender i.e 0.41/ μL (1.2/LPF). In normal urine hyaline cast could be found 0-2/LPF.

Path.cast are normally not present in urine. Results are 0.13/ μL (0.4/LPF) for female and 0.0/ μL(0.0/LPF). Reference values obtained for female were near zero.

DISCUSSIONS

Yeast are normally not present in urine however values obtained are near zero i.e 0.7/μL (0.1/HPF) in female and 0.2/μL (0.0/HPF) in male.

Mucus could be found in the urine of healthy persons. Mucus results are 0.78/μL (1.2/LPF) in female and 2.8/LPF in male.

Crystal and sperm were 0.0/μL or 0.0/HPF in female and male.

CONCLUSION In the present study, we have determined the reference range of urine

sediments from medical check up participants by using UF-4000

The results obtained could reflect the results of healthy population. The values obtained are useful for the analysis of wellness screening data.

The UF-4000 have new improvements which allows a better standardization of urine sediment analysis and it support in reporting quantitative results per field (HPF/LPF) and per μL unit.

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