Kridsada Sirisabhabhorn and Supaporn pumpa Clinical Microscopy & ParasitologyDivision, Department of Medical Technology Laboratory ,
ThammasatCharlermprakiethospital, Pathumtani, Thailand 12120
A Reliable and High Yielding Method for Isolation of Genomic DNA from Ammi MajusSandip Magdum
The developed protocol describes a cheaper, quicker and reliable method for the isolation of pure DNA from medicinal herbs, such as Ammi majus, which produces the secondary metabolites xanthotoxin and berganpectane having immense medicinal importance. Use of CTAB, liquid nitrogen and EDTA in different isolation protocols analyzed for A. majus, all were ended with polysaccharide and protein contamination with low purity of DNA (A260/280=1.3-1.6), revealed a need for method modification for the inexpensive and rapid isolation of pure DNA. Developed reliable and competent protocol isolated enough pure DNA (A260/280=1.81) without following time consuming lengthy steps and hazardous chemicals used in other protocols, which increase experimental costs, risk, and need expertise to perform. The explained protocol requires few chemicals and little time to obtain pure DNA having yield 688 μg/g of A. majus. A higher quantity of isolated DNA obtained from young fresh leaf samples than from either the callus or stem. A. majus is a pharmaceutically important medicinal herb, and the present protocol aids in the analysis and modification of its genes.
Rapid, cost effective and disposable device for saliva collection and process...Robert Gellibolian, Ph.D
Test and manufacture a device for rapid, cost
effective, non-invasive collection and ‘clean up’ of saliva (e.g.,
reduction of contaminants, matrix effects, etc) for early detection
of Malaria biomarkers (e.g., pfHRP2, etc). ‘Clean’ saliva offers
several advantages over blood; (1) No cultural / religious biases
associated with collection of blood, (2) reduced biohazard to
worker, (3) non-invasive, (4) easy
Bio-ingénierie hépatique / Bio-Printing du foie : Enjeux et espoirs
Jean-Charles Duclos-Vallée
Journées du Centre Hépato-Biliaire - JCHB 2019
Hépatologie
Benzonase® endonuclease: use and clearance with a TFF step in a cell culture-...Frédéric Sengler
Benzonase® endonuclease is a powerful tool for degrading all
forms of (deoxy)ribonucleic acids (DNA/RNA) in cell culture
harvests to base pair (BP) lengths under 8 units. It is effective
over a wide range of conditions including temperature, pH, and
varying concentrations of Mg2+, detergents, chelating agents,
and monovalent cations. It is often employed in the production
of viral vaccines, completely digesting DNA and RNA to improve
clearance and reduce solution viscosity.
Removing Benzonase® endonuclease, a 60 kD dimer, from the
process stream post-treatment may be achieved with Tangential
Flow Filtration (TFF) with the appropriate molecular weight
cut-off membrane (MWCO), typically 300 kD. This process has
heretofore not been well described in the literature, so the
present study fills this gap pairing Benzonase® endonuclease
treatment of a DNA-spiked inactivated flu virus cell culture
harvest with Pellicon® 2 cassettes, for clearance of the digested
DNA and remaining Benzonase® endonuclease by diafiltration.
A Reliable and High Yielding Method for Isolation of Genomic DNA from Ammi MajusSandip Magdum
The developed protocol describes a cheaper, quicker and reliable method for the isolation of pure DNA from medicinal herbs, such as Ammi majus, which produces the secondary metabolites xanthotoxin and berganpectane having immense medicinal importance. Use of CTAB, liquid nitrogen and EDTA in different isolation protocols analyzed for A. majus, all were ended with polysaccharide and protein contamination with low purity of DNA (A260/280=1.3-1.6), revealed a need for method modification for the inexpensive and rapid isolation of pure DNA. Developed reliable and competent protocol isolated enough pure DNA (A260/280=1.81) without following time consuming lengthy steps and hazardous chemicals used in other protocols, which increase experimental costs, risk, and need expertise to perform. The explained protocol requires few chemicals and little time to obtain pure DNA having yield 688 μg/g of A. majus. A higher quantity of isolated DNA obtained from young fresh leaf samples than from either the callus or stem. A. majus is a pharmaceutically important medicinal herb, and the present protocol aids in the analysis and modification of its genes.
Rapid, cost effective and disposable device for saliva collection and process...Robert Gellibolian, Ph.D
Test and manufacture a device for rapid, cost
effective, non-invasive collection and ‘clean up’ of saliva (e.g.,
reduction of contaminants, matrix effects, etc) for early detection
of Malaria biomarkers (e.g., pfHRP2, etc). ‘Clean’ saliva offers
several advantages over blood; (1) No cultural / religious biases
associated with collection of blood, (2) reduced biohazard to
worker, (3) non-invasive, (4) easy
Bio-ingénierie hépatique / Bio-Printing du foie : Enjeux et espoirs
Jean-Charles Duclos-Vallée
Journées du Centre Hépato-Biliaire - JCHB 2019
Hépatologie
Benzonase® endonuclease: use and clearance with a TFF step in a cell culture-...Frédéric Sengler
Benzonase® endonuclease is a powerful tool for degrading all
forms of (deoxy)ribonucleic acids (DNA/RNA) in cell culture
harvests to base pair (BP) lengths under 8 units. It is effective
over a wide range of conditions including temperature, pH, and
varying concentrations of Mg2+, detergents, chelating agents,
and monovalent cations. It is often employed in the production
of viral vaccines, completely digesting DNA and RNA to improve
clearance and reduce solution viscosity.
Removing Benzonase® endonuclease, a 60 kD dimer, from the
process stream post-treatment may be achieved with Tangential
Flow Filtration (TFF) with the appropriate molecular weight
cut-off membrane (MWCO), typically 300 kD. This process has
heretofore not been well described in the literature, so the
present study fills this gap pairing Benzonase® endonuclease
treatment of a DNA-spiked inactivated flu virus cell culture
harvest with Pellicon® 2 cassettes, for clearance of the digested
DNA and remaining Benzonase® endonuclease by diafiltration.
Polymerase Chain Reaction (PCR)-Based Sex Determination Using Unembalmed Huma...IOSR Journals
The strategy developed for sex determination in skeletal remains is to amplify the highly degraded DNA, by use of primers that span short DNA fragments. To determine sex of unembalmed human cadaveric skeletal fragments from Sokoto, North-western Nigeria, using Polymerase Chain Reaction (PCR). A single blind study of Polymerase Chain Reaction (PCR)-based sex determination using amelogenin gene and alphoid repeats primers on unembalmed human cadaveric skeletal fragments from Sokoto, North-western Nigeria, was undertaken. With amelogenin gene, genetic sex identification was achieved in four samples only. PCR Sensitivity = 40%, Specificity = 100%, Predictive value of positive test = 100%, Predictive value of negative test = 25%, False positive rate = 0%, False negative rate = 150%, Efficiency of test = 50%. Fisher’s exact probability test P = 1. Z-test: z-value = -1.0955, p>0.05; not statistically significant. With alphoid repeats primers, correct genetic sex identification was achieved in all the samples. PCR Sensitivity = 100%, Specificity = 0%, Predictive value of positive test = 100%, Predictive value of negative test = 0%, False positive rate = 0%, False negative rate = 0%, Efficiency of test = 100%. Fisher’s exact probability test P = 1. Z-test: z- and p values were invalid. The study, has demonstrated the applicability of PCR method of sex determination in unembalmed human skeletal fragments from Sokoto, Northwestern Nigeria. With amelogenin gene primers, correct genetic sex identification was achieved in four samples only. With alphoid repeats primers, correct genetic sex identification was achieved in all the samples. Therefore, alphoid repeats is more efficient and more reliable than amelogenin gene, in sex determination from unembalmed human skeletal fragments. This is the first known study determining the sex of unembalmed human skeletal fragments by means of PCR in Nigeria. There is need for further studies in Nigeria to complement the findings of this study.
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...journal ijrtem
Abstract: The present study addresses this issue by examining the patterning of Cytochrome Oxidase I diversity in the stone fish Uranoscopus oligolepis the structurally diverse group of Family Uranoscopidae. The sequences were analyzed for their species identification using BOLD’s identification engine. The COI sequences of U. oligolepis from different geographical regions were extracted from NCBI for intra species variation analysis. All sequences were aligned using Clustal W. The sequences were trimmed using software and phylogenetic tree was constructed with bootstrap test. The results showed that the cytosine content was high (31%). The least molar concentration was observed in guanine (19.5%) and Adenine (19.6%). Thymine was the second predominant in molar concentration next to thymine which is followed by adenine. The G+C content was found to be 49.6% and A+T content was 50.4%. Leucine and Alanine content was high in the amino acid composition. From the study it is assumed that the mitochondrial gene COI can be the potential barcoding region to identify an organism up to the species level. Keywords: COI, intra species, Uranoscopus oligolepis, barcoding, phylogenetic
Development of quality control assays for cell-based medicinal products (ISCT...Quality Assistance s.a.
Dr. Fabian Vandermeers from Quality Assistance spoke on Development of quality control assays for cell-based medicinal products at ISCT 2017 in London.
For more information on this topic, visit: http://www.quality-assistance.com/analytical-services/CBMPs
For more information on our expertise and services, visit: www.quality-assistance.com
Follow us on social media:
LinkedIn: https://www.linkedin.com/company/quality-assistance
Twitter: https://twitter.com/QA_Belgium
Facebook: https://www.facebook.com/QualityAssistanceBelgium
Google +: https://plus.google.com/103676189647965359292
Quality Assistance S.A. is a leading European Contract Research Organisation providing the pharmaceutical industry with all the analytical services required by EMA and FDA regulations for the development and marketing of innovative human medicinal products.
We assist our clients from candidate selection, through non-clinical and clinical studies, to marketing authorisation, using our state-of-the-art, product-dedicated expertise in analytical sciences.
For each customer and each project, we design customised solutions, define analytical protocols, develop and validate specific new analytical methods and perform characterisation, stability, pharmacokinetic, biomarker and immunogenicity studies as well as batch release testing, in order to evaluate the Quality, Safety and Efficacy of the given drugs.
Abstract book for the following conferences:
2013 2nd International Conference on Bioinformatics and Biomedical Science (ICBBS 2013)
2013 2nd International Conference on Environment, Energy and Biotechnology (ICEEB 2013)
2013 2nd International Conference on Chemical and Process Engineering (ICCPE 2013)
2013 2nd Journal Conference on Environmental Science and Development (JCESD 20132nd)
The conferences was held at Concorde Inn Kuala Lumpur International Airport, Malaysia on 09 June 2013
This presentation in mainly focused of understanding of automation and its utility in cytopathology. It will be very usefull for postgraduate in pathology, cytopathologist and cytotechnicians.
Clinical diagnosis of chronic myeloid leukemia by real time polymerase chain ...Teboho Mooko
Oncology study i did in my third year (2014). the study was basically about monitoring Chronic Myeloid leukemia (CML) using Real-Time PCR techniques to check how patients from Universitas Hospital responded to the treatment of Gleevec drug.
Polymerase Chain Reaction (PCR)-Based Sex Determination Using Unembalmed Huma...IOSR Journals
The strategy developed for sex determination in skeletal remains is to amplify the highly degraded DNA, by use of primers that span short DNA fragments. To determine sex of unembalmed human cadaveric skeletal fragments from Sokoto, North-western Nigeria, using Polymerase Chain Reaction (PCR). A single blind study of Polymerase Chain Reaction (PCR)-based sex determination using amelogenin gene and alphoid repeats primers on unembalmed human cadaveric skeletal fragments from Sokoto, North-western Nigeria, was undertaken. With amelogenin gene, genetic sex identification was achieved in four samples only. PCR Sensitivity = 40%, Specificity = 100%, Predictive value of positive test = 100%, Predictive value of negative test = 25%, False positive rate = 0%, False negative rate = 150%, Efficiency of test = 50%. Fisher’s exact probability test P = 1. Z-test: z-value = -1.0955, p>0.05; not statistically significant. With alphoid repeats primers, correct genetic sex identification was achieved in all the samples. PCR Sensitivity = 100%, Specificity = 0%, Predictive value of positive test = 100%, Predictive value of negative test = 0%, False positive rate = 0%, False negative rate = 0%, Efficiency of test = 100%. Fisher’s exact probability test P = 1. Z-test: z- and p values were invalid. The study, has demonstrated the applicability of PCR method of sex determination in unembalmed human skeletal fragments from Sokoto, Northwestern Nigeria. With amelogenin gene primers, correct genetic sex identification was achieved in four samples only. With alphoid repeats primers, correct genetic sex identification was achieved in all the samples. Therefore, alphoid repeats is more efficient and more reliable than amelogenin gene, in sex determination from unembalmed human skeletal fragments. This is the first known study determining the sex of unembalmed human skeletal fragments by means of PCR in Nigeria. There is need for further studies in Nigeria to complement the findings of this study.
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...journal ijrtem
Abstract: The present study addresses this issue by examining the patterning of Cytochrome Oxidase I diversity in the stone fish Uranoscopus oligolepis the structurally diverse group of Family Uranoscopidae. The sequences were analyzed for their species identification using BOLD’s identification engine. The COI sequences of U. oligolepis from different geographical regions were extracted from NCBI for intra species variation analysis. All sequences were aligned using Clustal W. The sequences were trimmed using software and phylogenetic tree was constructed with bootstrap test. The results showed that the cytosine content was high (31%). The least molar concentration was observed in guanine (19.5%) and Adenine (19.6%). Thymine was the second predominant in molar concentration next to thymine which is followed by adenine. The G+C content was found to be 49.6% and A+T content was 50.4%. Leucine and Alanine content was high in the amino acid composition. From the study it is assumed that the mitochondrial gene COI can be the potential barcoding region to identify an organism up to the species level. Keywords: COI, intra species, Uranoscopus oligolepis, barcoding, phylogenetic
Development of quality control assays for cell-based medicinal products (ISCT...Quality Assistance s.a.
Dr. Fabian Vandermeers from Quality Assistance spoke on Development of quality control assays for cell-based medicinal products at ISCT 2017 in London.
For more information on this topic, visit: http://www.quality-assistance.com/analytical-services/CBMPs
For more information on our expertise and services, visit: www.quality-assistance.com
Follow us on social media:
LinkedIn: https://www.linkedin.com/company/quality-assistance
Twitter: https://twitter.com/QA_Belgium
Facebook: https://www.facebook.com/QualityAssistanceBelgium
Google +: https://plus.google.com/103676189647965359292
Quality Assistance S.A. is a leading European Contract Research Organisation providing the pharmaceutical industry with all the analytical services required by EMA and FDA regulations for the development and marketing of innovative human medicinal products.
We assist our clients from candidate selection, through non-clinical and clinical studies, to marketing authorisation, using our state-of-the-art, product-dedicated expertise in analytical sciences.
For each customer and each project, we design customised solutions, define analytical protocols, develop and validate specific new analytical methods and perform characterisation, stability, pharmacokinetic, biomarker and immunogenicity studies as well as batch release testing, in order to evaluate the Quality, Safety and Efficacy of the given drugs.
Abstract book for the following conferences:
2013 2nd International Conference on Bioinformatics and Biomedical Science (ICBBS 2013)
2013 2nd International Conference on Environment, Energy and Biotechnology (ICEEB 2013)
2013 2nd International Conference on Chemical and Process Engineering (ICCPE 2013)
2013 2nd Journal Conference on Environmental Science and Development (JCESD 20132nd)
The conferences was held at Concorde Inn Kuala Lumpur International Airport, Malaysia on 09 June 2013
This presentation in mainly focused of understanding of automation and its utility in cytopathology. It will be very usefull for postgraduate in pathology, cytopathologist and cytotechnicians.
Clinical diagnosis of chronic myeloid leukemia by real time polymerase chain ...Teboho Mooko
Oncology study i did in my third year (2014). the study was basically about monitoring Chronic Myeloid leukemia (CML) using Real-Time PCR techniques to check how patients from Universitas Hospital responded to the treatment of Gleevec drug.
52.Iqbal J, Patil R, Khanna V, Tripathi A, Singh V, Munshi MAI, Tiwari R. Role of fractal analysis in detection of dysplasia in potentially malignant disorders. J Family Med Prim Care. 2020 May;9(5):2448-2453. doi: 10.4103/jfmpc.jfmpc_159_20. eCollection 2020 May. PubMed PMID: 32754518; PubMed Central PMCID: PMC7380790.
Liquid biopsy quality control – the importance of plasma quality, sample prep...Thermo Fisher Scientific
Liquid biopsy is emerging as a non-invasive companion to traditional solid tumor biopsies. As next generation sequencing (NGS) of circulating cell-free nucleic acids (cfNA = cfDNA and cfRNA) becomes common, it’s important to understand the impact of sample preparation on quality, specificity, and sensitivity of liquid biopsy tests. Plasma samples are often limited, and may have undesirable characteristics such as lipemia or hemolysis that contribute unwanted genomic DNA (gDNA) to the sample. Low cfDNA concentration can also limit the amount available for NGS library prep. In this study, we explore the effects of suboptimal plasma and low library input on liquid biopsy NGS, and discuss various techniques for in-process quality control of cfNA samples isolated from plasma
Kridsada Sirisabhabhorn, Supaporn Pumpa and Palakorn Puttaruk, Medical Techno...kridsada31
The comparative study of prevalence of human parasitic infection during 2014 –2015 year; the beginning year of approach to the AEC community.
Background:ParasiticinfectionsofhumanremainacauseofhealthproblemsinpartofSouthEastAsia(SEA) region.AccordingtoASEANEconomicCommunity(AEC) opening,pathogenicparasitescaneasilyspreadto10membercountriesbytravelers.
Objective:Theobjectiveinthisstudyweretoinvestigatehuman’spathogenicparasiteinfecalandbloodspecimensandcomparedprevalenceofinfectionbetween2014(beforeAECopening) and2015(AECcommunityyearopening) toinspectatrendofinfectionintoourcountry.
Materials/Methods:Datawerecollectedfrom9,608casestotalsinceJanuary2014toDecember2015.Stoolconcentrationtechniqueusingformalin‐ethylacetate,wasusedtoincreaseprobabilityoffindingparasitesinfeces.Ontheotherhand,thickandthinbloodsmearwerepreparedfromperipheralbloodsampleforinvestigatingblood‐borneparasiticinfection.Percentageofparasiticprevalenceineachyearwascalculatedandpresentedusingdescriptivestatistics.
Results:Theresultspresentedthattotalprevalencerateofparasiticinfectionswas1.24%(53/4,272) and1.42%(76/5,336) in2014and2015respectively.In2015,prevalencerateofBlastocystishominis (27/4,272) andStrongyloidesstercoralis (10/4,272) infectionsincreased1.4timeswhencomparedto2014(B.hominis (46/5,336)andS.stercoralis (16/5,336)).Inaddition,Opisthorchisviverrini (3/5,336) andMinuteegg(2/5,336) werenewemergedin2015aswellasotherparasitessuchasGiardialamblia,Taeniaspp.,EntamoebacoliandPlasmodiumspp.,buttheywerefoundinthelowinfectionrate.
Conclusions:Thisstudyrevealedthattotalparasiticinfectionratein2015wasslightlyincreasedfrom2014.OpeningofAECseemstoaffectpublichygieneprobleminthisregion.Thisstudyserveddatabaseformonitoringandpredictingthetrendofprevalencerateinthefuture.Nevertheless,continuousdatacollectionwillfulfillandimprovetherecentinformation.
Keywords:AEC,Parasite,Infection,Trend,Prevalence
Increase in the prevalence of Clostridium difficileproducing toxin B in fecal...kridsada31
Kridsada Sirisabhabhorn1,2, Wanna Chaijaroenkul2, and Kesara Na-Bangchang2*
Abstract: Toxin A (TcdA) and B (TcdB) are virulence factors of Clostridium difficilethat cause diarrhea and pseudomembranous colitis. The global prevalence of C. difficileinfection (CDI)has been increasing due to resistance of the bacteria to the standard treatment with vancomycin. The aim of the study was to investigate the prevalence rates ofC. difficileproducing TcdAand TcdBin fecal specimens collected in 2014 (n=646) and 2015 (n=649) from CDI patients at ThammasatChalermprakietHospital, Pathumtani, Thailand. TcdAand TcdBin all samples were detected using immunochromatographicdipstick test. The number (%) of bacteria producing TcdA, TcdB, and a mixture of TcdAand TcdBfound in 2014 were 11 (1.7%), 14 (2.2%), and 22 (3.41%) samples, respectively. The corresponding numbers (%) for samples collected in 2015 were 5 (0.7%), 27 (4.2%), and 17 (2.6%), respectively. The prevalence of fecal specimens with TcdBpositive was significantly increased in 2015 compared with 2014. This observation may be a warning sign for the progress of C. difficileresistant strains. Close monitoring of their prevalence rates in the hospital including sensitivity to chemotherapeutics is essential.
Key words: C. difficile, TcdA, TcdB, prepvalence, resistance
Thecomparison methods of amphetamines detection in urine samples in Thammasat...kridsada31
Thecomparison methods of amphetamines detection in urine samples in ThammasatUniversity hospital, Pathumtaniprovince.
KridsadaSirisabhabhornSupapornPumpaNarumonSereekhajornjaruand PalakornPuttarak
Department of Medical Technology Laboratory, ThammasatUniversity Hospital Pathumtani, Thailand
Comparison and Evaluation of FUS-2000 Urinalysis Hybrid and Conventional Micr...kridsada31
Comparison and Evaluation of FUS-2000 Urinalysis Hybrid and Conventional Microscopic Examination
PrangthipSukboon1, SupichchaDuangson1, KridsadaSirisabhabhorn2,Chollanot Kaset1, SupapornPumpa2, DaungnatePipatsatitpong1*
1Department of Medical Technology, Faculty of Allied Health Sciences, ThammasatUniversity2Division of Clinical Microscopy and Parasitology, Department of Medical Technology Laboratory, ThammasatUniversity Hospital
Malaria situation in Thammasat Charlerm Prakiet hospital Pathumthani/Kridsada31kridsada31
Pumpa S.1, Sirisabhabhorn K.1, Cheoymang A.2, Sintana T1. and Puttarak P1.
1Department of Medical Technology Laboratory, Thammasat Charlerm Prakiet hospital Pathumthani
2Pharmacology unit faculty of Allied Health Sciences Thammasat University Pathumthani
Comparison of reticulocyte count by VCS technology; Beckman Coulter Hematolog...kridsada31
Kridsada Sirisabhabhorn1 Anurak Choeymang2 Supaporn Pumpa1 Naiyana poottasane1 and Palakorn Puttaruk1
1Department of Medical Technology Laboratory, Thammasat University hospital, Pathumthani, Thailand
2Pharmacology and Toxicology Unit, Faculty of Allied Health Sciences, Thammasat University, Pathumthani, Thailand
Trend of parasitic infection rates of Thammasat University Hospital’s patient...kridsada31
Kridsada Sirisabhabhorn Supaporn Pumpa Krim Kamyod and Palakorn Puttaruk Microscopy Unit, Department of Medical Technology Laboratory, Thammasat University Hospital, Pathumthani, Thailand
Improvement of bacterial estimation in urinalysis by iQ®200 ELITE®: capture i...kridsada31
Kridsada Sirisabhabhorn1 Anurak Choeymang2 Supaporn Pumpa1 Krim Kamyod1 and Palakorn Puttaruk1
1Department of Medical Technology Laboratory, Thammasat University hospital, Pathumthani, Thailand
2Pharmacology and Toxicology Unit, Faculty of Allied Health Sciences, Thammasat University, Pathumthani, Thailand
Improvement of bacterial estimation in urinalysis by iQ®200 ELITE®: capture i...
The introduction of well trained experience with iQ®200i microscopic analyzer improved/Kridsada31
1. Kridsada Sirisabhabhorn and Supaporn Pumpa
Dysmorphic rbc (dRBC) is leaked morphologic of red blood cell which infiltrated
throughout from pathologic of glomerular in kidney. The variant types of dRBC morphology
are significant for indication of glomerular bleeding and grading of severe glomerular
disease. Thus, D1 cell is like a ring-like shape and severe cytoplasmic color loss with
membranous protrusions or blebs, D2 is like a doughnut-like shape and moderate
cytoplasmic color loss with membranous protrusions or blebs. and D3 cell is like a doughnut
shape and mild cytoplasmic color loss without membranous protrusions or blebs.(1) In the
present, various choices of automated analyzer technology could be support technician
such as reduce TAT (Turn Around Time) and reduce workload like as principle capture
image.(3) and flow cytometry. Nevertheless advances in skills and experience is still
important for classification of pathological urine element, especially dRBC that notified
glomerular disease exclude non-glomerular disease.
Objectives
1. To study performance of 2 individuals who are well trained and non
trained in order to screen abnormal cases and identify dRBC and urine
elements with clinical specific kidney diseases from iQ®200i microscopic
automated analyzer.
2. To demonstrate achievement of the well trained person to classify
morphology of dRBC.
Material & Method
Total 143 cases of urinalysis were 74 male and 59 female cases that
received from IPD patients of Thammasat Charlermprakite hospital,
Pathumtani, Thailand. All samples were analyzed by 2 technicians (well
trained and non-trained in dRBC identification) using iQ®200i. Over 3 ml
and uncentrifuge of midstream urine permitted for input to iQ®200i
system; combine chemistry and microscopic part. The technical
reclassified elements on desktop of iQ®200i, the principle of capture
image urine elements. Three major categories which are red blood cells,
white blood cells and squamous epithelial cells were recognized by
analyzer. On the contrary, minor categories depended on human
verification. During the experiment if previous specimen was high
concentration of cells later introduced to clean with cleanser and diluents
for eliminated carry over. dRBC in all cases were confirmed by
centrifugation at 400 x g for 5 min. and subjected to brightfield
microscopic examination. Finally, the results were analyzed by descriptive
statistic.
Results
Table 1 and Bar chart 1. shown that the efficiency skill in urine elements screening between
non-trained with well trained person. After urine elements were easily screened by iQ®200i,
there were abnormal 49 cases which needed confirmation by brightfield microscopic
examination as shown in table 2. The results were demonstrated that 10 cases of dRBC in
variant classification of dRBC as D1, D2 and D3 cell and mixed erythrocytes morphology
respectively as on table 3.
Classification
of personal skill
All cases (143)
Released
cases
% of all cases
Non-trained 57 39.86
Well trained 94 65.73
10 cases of dysmorphic red blood cell
Cell types no. of cases % of all cases
D1 0 0
D2 6 4.20
D2 +D1 1 0.70
D2 +D3 1 0.70
D3 2 1.400
10
20
30
40
50
60
70
80
90
100
Non‐trained Well trained
% screening cases achievement of total cases (143)
Non‐trained
Well trained
Discussion & Conclusion
Introduction
Abstract
Class of personal
skill
Appearance cases
of dRBC
After centrifugation
Non-trained
(cases)
Well trained
(cases)
dRBC found 4 10
dRBC not found 139 133
References 1. Nagahama, D.,Yoshiko, K., Morita, M.W.Y., Iwatani, Y. and Matsuo, S. 2005. A useful new classification of dysmorphic urinary erythrocytes. Clin Exp Nephrol 9:304–9.
2. Crop., M.J., de Rijke., Y.B., Verhagen., P.C.M.S., Cransberg, K.and Zietse, R. 2010. Diagnostic Value of Urinary Dysmorphic Erythrocytes in Clinical Practice. Nephron Clin Pract ;115:c203–c2.
3. Sarvary, E., Lee, D., Varadi, J. Varga, M., Gaal, I. and Chmel, R. et. al, 2010. The iQ200 Microscopic Analyzer is valuable tool for evaluation of urinary sediment at transplanted patients. Interventional Medicine & Applied
Science, Vol. 2 (1). 22–6.
This study demonstrated that experience from well trained person for diagnostic urine element is important in routine urinalysis. In the well trained personal
determined easy elements on monitor of iQ®200i who approved screening raise up to 25.87% of all cases more than non-trained personal. This experiment revealed that
there were 10 dRBC cases from 49 abnormal cases. Moreover, variant morphology of dRBC in this study were described as D2, D3 cell sand mixed type D2 with D1
cells and D2 with D3 cells. This study is preliminary survey in significant of well trained experience in iQ®200i usual which influence to diagnosis dRBC erythrocyte that
associated with glomerular disease. Most kinds of dRBC morphology were appeared as D2 cell [moderate glomerular damage] below as D3 cell [mild glomerular
damage]. Particularly, it is difficult for D3 cell investigation because it may be missed in diagnosis by less experience person. However,D3 cell is an early marker for
indication of glomerular disease.(2) This preliminary study will be create different experience field in usual automate analyzer effect to diagnosis abnormal urine elements.
The sensitivity, specificity and PPV in kidney disease should be further study. The benefit of this study is to enhance personal performance in order to identify dRBC and
increase quality in routine urinalysis.
Clinical Microscopy & Parasitology Division, Department of Medical Technology Laboratory ,
Thammasat Charlermprakiet hospital, Pathumtani, Thailand 12120
Key words : Dysmorphic rbc, iQ®200i, glomerular disease
Classification of dysmorphic erythrocyte (dRBC) plays a crucial role in indication of glomerular damage. Moreover, variant type of morphology dRBC is used to
demonstrate the severity of glomerular disease. Although, the iQ®200i microscopic capture image analyzer can be used to reduce laborious and time consuming, the well
trained of device usual is still improving for dRBC investigation. The aim of this study is to evaluate the efficiency of 2 individuals of well trained and non-trained using
with the iQ®200i for improving the dRBC diagnosis and rule out normal urine before dRBC confirmation by microscopic. One hundred and forty-three urine samples of
IPD patients in Thammasat Charlermprakiet hospital, Pathumtani, Thailand were collected and subjected to urinalysis by iQ®200i analyzer and 2 medical technologists.
Next, abnormal cases were centrifuged at 400x g 5 min and then the pellet was observed under bright-field microscope. The urinalysis results between 2 individuals were
compared by using descriptive statistic. The results were demonstrated that the well trained performed raise 25.87% of cases in urinary screening so that better than
non-trained skill. Especially, classification of dRBC were described as D2, D3 cell and mixed types of D2+D1 cells and D2+D3 cells as 4.2, 1.4, 0.7 and 0.7%,
respectively. The advantages of this study is to emphasize in dRBC diagnosis and activate personal skill for usual device. However, sensitivity, specificity and PPV of
predicted kidney disease should be further study.
The introduction of well trained experience with iQ®200i microscopic analyzer improved
dysmorphic erythrocyte diagnosis
Table 1.
Bar chart 1.
Table 2.
Table 3.
D2+D1 cells D2+D3 cells
Acknowledgements : We would like to thank for her kindness of Lecture Ornrudee Khantisitthiporn, Faculty of Allied Health Sciences, Thammasat University for approve literature and Mr.Palakorn Puttarak, Lab director of Medical Technology
Laboratory, Thammasat Charlermprakiet Hospital for support poster presentation.
D1 cell
D3 cell
D2 cell
Photo by Kridsada Sirisabhabhorn, MT.