Modern Medical Laboratory Journal

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Background and Objective: Shigella species belong to the family Enterobacteriaceae, which cause dysentery with abdominal pain and tenesmus in human. Our country is one of the endemic and occasional epidemic areas of the disease. In the present study, Repetitive Extragenic Palindromic Polymerase Chain Reaction (REP-PCR) technique that has high differentiation and specificity power compared to phenotypic markers, was used to investigate diversity and geographical distribution of colones as well as typing of Shigella strains.

Materials and Methods: In this study, a total of 40 Shigella sonnei samples were isolated from stool of patients with diarrhea and divided into 5 groups. After identification and confirmation of the isolates by biochemical and serotyping methods, one colony of each isolate was cultured on LB medium and after DNA extraction, PCR was performed. After electrophoresis of the PCR product, gel images were saved in the computer for further analysis and typing of the isolates. 

Results: In this research, 40 isolates were examined using REP-PCR and a similarity matrix was constructed based on Dice's coefficient. According to this matrix, some isolates were completely similar (genetic similarity coefficient= 1) and some isolates showed the least similarity (genetic similarity coefficient= 0). The dendrogram was obtained using the UPGMA algorithm. The calculated Cophenetic correlation coefficient for this dendrogram was 0.91645.

Conclusion: From the results of the present study, it was concluded that we can type the Shigella sonnei strains using palindromic repetitive sequences. The extent of polymorphism indicates that REP-PCR technique is a useful method for genetic variation analysis in molecular typing of shigella sonnei strains. Although, some strains were completely similar, a high genetic variation was found among the studied population of Shigella sonnei. This level of variation is probably due to wide geographic distribution of this species in Iran.

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Background and Objective: Acinetobacter is a genus of non-fermenting Gram-negative cocci or coccobacilli, which have low nutritional requirements for growth and can survive for a long time in adverse conditions, on dry surfaces, and also in aqueous environments. The importance of the members of Acinetobacter genus as pathogens involved in nosocomial infections, is increasing. Acinetobacter baumannii is the most common species involved in a broad spectrum of nosocomial infections, including pneumonia, bacteremia, surgical wound infections, urinary tract infections, and meningitis. In this study, enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR) technique was used for analysis and molecular typing of Acinetobacter strains, which has high discrimination power compared to phenotypic markers. 

Methods: In the present study, a total of 40 A. baumanniies strains were isolated from patients hospitalized in Tehran hospitals. After identification and confirmation of the isolates by serotyping and biochemical tests, a single colony of each isolate was cultured on liquid LB medium, and after DNA extraction, PCR was performed. After electrophoresis of PCR product, gel images were stored electronically for analysis and comparison of the isolates. 

Results: In this study, 40 strains of A. baumannii were analyzed by ERIC-PCR method, of which 29 strains were typed into 10 groups and 11 other strains had no PCR bands or had a band that could not be assigned to any of the above groups.

Conclusion:  In this study, it was found that A. baumanniie strains could be typed using repetitive sequences. This extent of polymorphism shows that ERIC-PCR is a useful method for analysis of genetic variation of A. baumannii strains. High genetic variation of A. baumannii strains may be due to wide geographical distribution of this species in Iran 

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Background and Objective: Beta-2 Microglobulin (β2M) is a middle molecular weight uremic toxin. Lack of β2M removal leads to β2M accumulation and amyloidosis in hemodialysis patients. The present research aims to determine the relationship between β2M and inflammatory factors, such as C-reactive protein (CRP), albumin, and high-density lipoprotein (HDL) in hemodialysis patients. 

Methods: Fifty-four hemodialysis patients were selected and their pre- and post-dialysis serum levels of β2M, CRP, albumin, and HDL, were measured.

Results: There was an obvious inverse relationship between β2M level and serum level of albumin, while no relationship was found between β2M and CRP or HDL.

Conclusion: According to the findings of this study, it was identified that inflammatory-induced increase in β2M level in dialysis patients leads to reduced serum albumin

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Background and Objective: Use of elastin-like proteins (ELPs) provides high-performance protein purification without need for chromatography. In line with cost reduction and facilitation of recombinant proteins purification, which represent a high percentage of production costs, in this project, we eliminated the need for proteases in the process of separation of recombinant proteins from ELP by designing a cassette using ELPs properties as well as insertion of autocatalytic intein protein between the recombinant protein and ELP.
Methods: In this study, at first Mxe GyrA intein gene was amplified from pTXB1 vector by PCR method and cloned into pUC57-hEGF vector. Then, 8xELP repetitive sequences were first cloned in pUC57 vector and then into pUC57-intein-hEGF vector in the upstream of Intein-hEGF.
Result: The design and construction stages of pUC57-8xELP-Intein-hEGF cassette was successful and the accuracy of 8xELP-Intein-hEGF was confirmed by sequencing.
Conclusion: The use of ELP-intein cassette provides recombinant protein purification only with steps consisting of temperature, salt, and centrifugation, without need for proteolytic enzymes, and access to this technology provides the possibility of production and purification of recombinant proteins with minimum cost and facilities. 

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Backround: The present study investigated the correlation between p53 gene codon 72 polymorphism and 6 other genetic single nucleotide polymorphisms (SNPs) in patients with cervical cancer infected by HPV.

Methods: 450 patients with cervical cancer (280 Squamous cell carcinoma and 170 Adenocarcinoma) were followed at hospitals in Iran from Dec. 2014 to Apr. 2015. Moreover, 100 age/sex-matched were used as the control group. HPV was detected by LINEAR ARRAY® HPV Genotyping Test. Allelic frequency of 6 gene polymorphisms was detected by the amplification-refractory mutation system (ARMS).

Results: From 450 patients, 408 cases (90.66%) were positive for HPV. Four genotypes were observed as single infections (16, 18, 31, and 45). The most common genotypes were HPV-16 (73.52%), HPV-18 (23.28%), HPV-31 and 45 (3.17%), respectively. 306 samples were arginine-arginine homozygous (70.6% and 71.4% of adenocarcinoma and squamous cell carcinoma, respectively), 70 cases were arginine-proline heterozygous (17.6% of adenocarcinoma and 23.8% of squamous cell carcinoma), and 20 cases were as proline-proline homozygous (11.8% and 4.8% of adenocarcinoma and squamous cell carcinoma, respectively).

Conclusion: The prevalence of HPV was 84% and that was the estimation of the Global Burden among Iranian patients with cervical cancer (85% - 99%). There was no correlation between mutations in the p53 allele and the size/type of tumors, while we found a correlation between mutations in p53 alleles and age. Therefore, XRCC1 G399A SNP and TP53 G72C SNP were significantly correlated with the cervical cancer.

Abbreviations:

HPV: Human papillomavirus; SNP: Single nucleotide polymorphism; CDKN1A: cycling-dependent kinase inhibitor 1A; TP53: Tumor protein p53 (also known as p53); P53c72: p53 gene codon 72; ATM: Ataxia-telangiectasia mutated; HDM2: Human double minutes 2 (also known as MDM2); LIG4: DNA ligase IV; XRCC1: X-ray repair cross-complementing 1; XRCC3: X-ray repair cross-complementing 3; TGFB1: Transforming Growth Factor Beta 1; HWE’: Hardy-Weinberg equilibrium; FIGO stage: International Federation of Gynecology and Obstetrics tumor stage; 

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Regenerative medicine, deals with functional reconstruction of damaged tissues or organs after severe injuries chronic diseases, while body's natural responses are not sufficient. In this field, stem cells due to their exclusive potential in self-renewal and differentiation into other cell types, are the main sources of functional cells in regenerative medicine. However, challenges in stem cell culturing highlighted the need for new methods, which in addition to maintaining cell viability and functionality, can control the precise microarchitecture for cells in a three dimensional structure. In this review we focus on the application of different types of bioprinting technology in regenerative medicine and we overview how this method has been able to make progress in 3D-cell culturing and tissue engineering protocols.

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Introduction: The aim of this work is to present the results of a comparative study between the determination of glycated hemoglobin (HBA1C) on the ADAMS A1c® (ARKRAY) and Capillarys 2 Flex Piercing® (SEBIA).
Materials and methods: 310 venous blood samples were randomly selected from routine HBA1C tests. The HBA1C assay was performed on ADAMS A1c® (ARKRAY) and Capillarys 2 Flex Piercing® (SEBIA) during the same day. The data obtained were analyzed by the statistical software MedCalc Version 15.1.0.
Results: The results obtained show a good correlation between the 2 methods: the equation of the Passing-Bablok line is of type Y (Capillarys 2 Flex Piercing®) = -0.550 + 1.119 X (ADAMS A1c) the 95% confidence interval of this slope is -0.6467 to - 0.4414 with r = 0.982 and p <0.0001. The Bland-Altman plot shows that the average bias between the two methods is in the order of 0.3 and that the difference between the Capillarys and HPLC measurements of Hba1c is in the range of +1.96 to -1.96 and the Deming regression equation Y (Capillarys 2 Flex Piercing®) = -0.3388 + 1.0911 X (ADAMS A1c).
Conclusion: Our study shows a good agreement of HBA1C results between the Capillarys 2 Flex Piercing® (SEBIA) and ADAMS A1c® (ARKRAY). Laboratory work requires professionals to take into account variations in results when changing methods in their routine work by comparing their results on the various methods.

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Objective: This study aims to estimate and compare the time-course change in blood glucose levels by blood cells in serum, and plasma with or without preservatives, which may reflect the rate of glucose utilization by blood cells.
Method: This laboratory-based cross-sectional study was carried out using a blood specimen of 28 participants among which 14 were diagnosed with diabetes and 14 were non-diabetic. Fasting blood specimen was collected in a plain tube, Ethylene Diamine Tetra Acetic Acid (EDTA) tube, and EDTA+ Sodium Fluoride (NaF) tube. The test was performed by hourly estimation of glucose for 24 hours. Time-course changes in glucose levels in serum and plasma with or without NaF preservative were statistically compared using ANOVA test.
Result: Serum and EDTA plasma glucose levels decreased gradually after the 3rd hour to 24th hour in comparison to EDTA+NaF plasma (p<0.05).  The rate of glucose utilization by blood cells was significantly higher in clotted blood and anticoagulated blood (EDTA) specimens in comparison with anticoagulated blood (EDTA) containing preservative (NaF) ((p<0.05). In addition, decreased rate of glucose utilization was observed in hyperglycemic specimens compared to that of normoglycemic blood.
Conclusion: Higher rate of glucose utilization by blood cells observed in serum and EDTA plasma represents a pre-analytical error in a long-standing specimen. The use of preservative NaF with EDTA significantly prevents cellular glucose utilization and stabilize plasma glucose level.  In contrast, this study also shows further insight into the reduced cellular metabolic rate of glucose utilization in diabetes mellitus.

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Introduction: The verification/validation of analytical equipment and methods is both part of this reasoning and an indispensable condition for their use and is one of the priorities of the medical biologist. The aim of our study is to verify the electrophoresis of serum proteins on the Cappilarys 2 Flex Piercing automaton.
Materials and methods: The evaluation methodology concerned the scope A which is based on the recommendations of the Valtec protocol of the French Society of Clinical Biology, as well as those of the SH-GTA O4 protocol of the COFRAC (Comité français d’accréditation). We studied the repeatability on normal and pathological serum samples, and the reproducibility on normal and pathological internal quality control samples.
Results: The values of the coefficient of variation of repeatability and reproducibility obtained by our study for each serum protein fraction (Albumin, Alpha-1, Alpha-2, Beta-1, Beta-2, Gamma globulins), are overall satisfactory and are in accordance with the requirements issued by the supplier and those issued by RICOS. In addition, these results are consistent with those of other similar studies.
Discussion and conclusion: This type of study will provide a solid basis for the realization of an accreditation procedure for the tests used in our laboratory. For any laboratory wishing to be accredited according to the ISO 15189 standard, the validation/verification of methods is a determining criterion. It is an essential step to be taken before the implementation of the newly acquired equipment.

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Objective: The study aimed to evaluate the performance of the liver profile during both the pre-analytical and analytical phases, focusing on Sigma metrics and the Quality Goal Index (QGI) at Benghazi Medical Centre, a Libyan.
Method: Data were gathered through a questionnaire focusing on pre-analysis, including request form eligibility and sample quality. The study assessed the quality of 200 request forms and 12,256 samples by computing Sigma levels and defects per million opportunities (DPMO). Additionally, the study analyzed three months of records for both normal and pathological Internal Quality Control (IQC) levels. For IQC performance evaluation, Sigma metrics and (QGIs) were calculated.
Result: The study uncovered a high frequency of pre-analytical errors in laboratory processes. These errors primarily revolved around deficiencies in request forms, leading to 34.6% being deemed ineligible, contributing to an overall low sigma level of 1.99.
Sample quality was also a concern, as 30% of samples suffered from insufficient quantity, resulting in a suboptimal overall sigma level of 2.91. Notably, total protein (T.P.) at level I exhibited excellent performance with a remarkable sigma value of 5.57. Additionally, three parameters, including T.P. at level II, AST at level II, and ALT at level II, demonstrated good performance with sigma levels of 4.40, 4.37, and 4.03, respectively.
Conclusion: We found that the performance of most parameters, excluding those mentioned, ranged from marginally acceptable to unacceptably low. Addressing these issues is crucial for enhancing diagnostic procedures' overall quality and reliability in healthcare settings.

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Background and Objective: The measurement of the coefficient of variation (CV%) and bias has been used to check the precision and accuracy of laboratory-generated data. Indices like the capability index (Cp and Cpk) and the quality goal index ratio (QGI) take into account both CV and bias in order to categorize the results of various parameters, allowing for necessary actions to improve the outcomes of poorly performing parameters.
Materials and Methods: This is a retrospective study, and data required for the study were extracted between January 2024 and June 2024 from a tertiary care government hospital in Delhi. The Cp, Cpk, and QGI were calculated, and six-month averages of these indices were taken to categorize the parameters.
Results: For level 1 of internal quality control (IQC), the Cp indicated that sodium, chloride, urea, and low-density lipoprotein (LDL) performed marginally well, while potassium performed poorly. Meanwhile, the Cpk showed poor performance for sodium, potassium, chloride, urea, LDL, amylase, and lactate dehydrogenase (LDH). Sodium, potassium, chloride, urea, and creatinine had a QGI of <0.8, indicating that the bias was due to imprecision, while LDL had a QGI of >1.2, indicating that the bias was due to inaccuracy. For level 2 of IQC, Cp showed that sodium, chloride, LDH, and LDL were in the marginal range, and potassium performed poorly, while Cpk indicated poor performance for sodium, potassium, chloride, urea, protein, LDL, and LDH. Sodium, potassium, chloride, urea, and protein had a QGI of <0.8, indicating that the bias was due to imprecision, while the QGI of LDL was between 0.8 and 1.2, indicating that the bias was due to both inaccuracy and imprecision.
Conclusion: This study shows that Cp, Cpk, and QGI can be used to assess the analytical performance of a clinical chemistry laboratory and to implement measures for improving quality assurance.