Medical Laboratory Journal

Comparing MALDI-ToFMass Spectrometry with Molecular and Biochemical Methods in Identifying Enterococcus Faecium and Enterococcus FaecalisIsolated from Clinical Samples

m MohebatiMobarez

Abstract Background and Objective: Enterococci are Gram-positive members of human gastrointestinal flora,in Dairy products and environment. they have emerged as important causes of opportunistic nosocomial infections in recent years. In this study we aimed to investigat and compare the efficiency of MALDI-TOFmass spectroscopy method through Biochemical and Molecular methods for detecting Enterococcus faecalis and Enterococcus faecium. Materials and Methods:seventhy five clinical samples were collected for biochemical, molecular and mass spectroscopy investigations. Samples were treated with Esculin hydrolysis, Catalase, Pyrrolidonylaminopeptidase, 6.5% NaCl solution, motility, 0.04% Tellurite, L-Arabinose and Sorbitol. Using specific primesallele specific PCR was used.The samples were then analyzed by MALDI-TOF mass spectroscopy and Biotyper 3 software. Results:Enterococcus faecium andEnterococcus faecaliswere detected in thirty and forty two samples, respectively whereas three samples showed both bacterial infections. Using biochemical analysis, two E.faecium isolates were Arabinose negative and one E. faecalis isolates was Telliurite negative. All sampleswere showed correct bands in PCR results but twoof them didn't show clear bands(on agarose gel). In mass spectroscopy analysis all strains were correctly detected and well defined. Conclusion: According to our results, MALDI-TOF mass spectrometry in comparison with Molecular and Biochemical Methodscould be a reliable and accurate method that can easily and quickly identify and differentiate Enterococcus faecium and Enterococcus faecalisin clinical samples. Key words:Enterococcus faecalis, Enterococcus faecium, MALDI-TOFmass spectrometry,PCR

The Comparison of Serum Vitamins A and E in Irritable Bowel Syndrome patients with Control Group

HR Joshaghani

Abstract Background and objectives: Irritable bowel syndrome (IBS) is one of the most common gastrointestinal diseases. It is a functional bowel disorder characterized by chronic abdominal pain and alternation of bowel habits with no structural abnormality. The prevalence of this disease was estimated 10-20 percent in the world. However, the cause of IBS is still unknown. Regarding to intestinal absorption disorder of nutrients, the malabsorption of vitamin may occur. The aim of this study is to evaluate serum levels of vitamins A and E in IBS patients. Materials & Methods: This case - control study was done on 94 patients whom their IBS disease were confirmed by a gastroenterologist in Golestan province. The control group was selected from healthy people, who didn't show any signs of digestive problems in past two years. The age and sex were matched with the cases group. Using HPLC method, Fasting blood samples were collected .Followed by measurement of Serum levels of vitamins A and E. Results: the meanserum levels of vitamin A and E in patients and control group were 57.0 ± 114.8μg / dl and23.8 ± 55.9 μg / dl, and 0.50 ± 0.24 mg / dl and 1.93 ± 1.86 mg / dl, respectively. (P <0.05). in men, the deficiency of Vitamin A and E, were7.70 and 7.10 percent and 6.76 and 3.7 in women. (P <0.05).In general, 1.1percent of IBS patients showed Vitamin A deficiency and 93.6 percent of them had vitamin E deficiency (P <0.05). Conclusion: In this study, a significant decrease in vitamin E levels was observed in patients with IBS. Due to antioxidant activity of vitamin E, the deficiency of this Vitamin, can increase the oxidative factors leading to intestinal damages and it is expected to decrease the amount of vitamins, subsequently Keywords: Irritable Bowel Syndrome, Vitamin A, Vitamin E, Gorgan

Evaluation of serum magnesium in type II diabetic patients

F Niknazar

Abstract Background and Objective: Some studies show that, there is a relationship between decrease of serum magnesium (Mg) level and the insulin resistance in diabetic type II patients. The aim of this study was to compare the Mg level in in type II diabetic with non-diabetic individuals. Materials and Methods: Diabetic patients(n=35) and control group (n=35),<35 years old, were randomly selected from the hospital of Imam Ali, Andimeshk. To compare diabetic patients and control group, the t-student statistical method and SPSS software, were used. Results: Mg levels in diabetic and non diabetic groups were 1.33±0.27 and 1.62±0.21 mg/dl, respectively, and the difference was statistically significant (p=0.0000014). A reversed significant relationship was seen between FPG and Mg among type II diabetic patients, whereas no significant relationship was seen between Mg and other variables, (TG, Chol and HDL). In non-diabetics group, there was a reversed significant relationship between the Mg and Chol, Whereas, no relationship was seen between other variables and magnesium. Conclusion: In diabetic patients, with increased level of FPG, serum Mg levels were decreased significantly. Moreover, serum Mg levels in diabetic patients were significantly lower than non-diabetics. Keywords: Magnesium, Diabetes type 2, Cholesterol, Triglyceride, High density lipoprotein

The Rate of HEP-2 Cellular Invasion of Salmonella Serogroups in Diarrhea Patients

MM Soltan Dallal

Abstract Background and Objectives: gasterointertidis is one of the most common forms of Salmonellosis, which is a worldwide problem. The invasive characteristic of intestinal bacteria is one of their pathogenicity Mechanisms , which can be easily investigated by cell culture technique. In this study ,the invasive characteristic of some Salmonella serogroup were investigated by using HEP-2 cell. Methods and Material: The rectals soap were prepared from 280 diarrhea patients referred to Imam Khomeyni and children medical centres , 140 with bloody diarrhea and 140 with watery diarrhea as a comparison group. The rectal soap was taken before patients taking any antibiotics, and 140 rectal specimens were taken from healthy people as a control group. All the samples were inoculated in differential and selective media, like Hektoen enteric agar and Xylose lysine deoxycholate (XLD) agar .After incubation at 37C for 24 hours, the colonies were examined and identified by conventional biochemical and serological tests. Using HEP-2, cellular invasion characteristic of Salmonella serogroups was assessed. Moreover, the antibiotic resistance patterns were performed according to Clinical and Laboratory Standards Institute (CLSI). Results: Of all tested samples, 35(8.3%) are Salmonella strains. The frequency of Salmonella is reported for bloody diarrhea (5.2%) , watery diarrhea ( 1.7%) and control group( 1.4%) .The most abundant serogroups with invasive characteristic, using HEP-2 cell culture, are serogroup B ( 62.9%) and D (17.2%). Conclusion The results obtained in this study show that the majority of Salmonella isolates are without invasive characteristic. Key words: Salmonella, Diarrhea, Cell invasion, Cell culture

Isolation of Lactobacilli from Dairy Products and Their Effects on the Main Pathogenic Bacteria in Stomach and Intestine

K Dorri

Abstract Background and objective: The effects of lactobacillus Species and their metabolites on dairy products were proved as probiotics on pathogenic bacteria. The aim of this study was to isolate the lactobacillus from dairy product (Yogurt, cheese, milk and whey …) and to determine the antimicrobial activity against some pathogenic bacteria (Salmonella typhimurium, E.coli, Helicobacter pylori. Material and methods: Samples of dairy products were collected and were serially diluted in peptone medium. To isolate lactobacillus, diluted sample were plated on MRS Agar medium and incubated at 37ºc for 48-72h in anaerobic condition. Identification of the lactobacilli was performed according to their morphological, cultural, physiological and biochemical characteristics. The inhibitory effect of isolated lactobacillus strains were performed against pathogenic bacteria by Agar Well Diffusion Assay and Disc method. Results: of 50 samples of isolated Lactobacillus from dairy products (including 11 identified Lactobacillus), 19 samples have inhibitory effect on pathogenic bacteria such as Salmonella typhimurium, Escherichia coli and Helicobacter pylori. The most identified strains areLactobacillusbulgaris, Lactobacillus Salivarius and Lactobacillus delbrueki. Conclusion: According to this study, some lactobacillus strains isolated from dairy products are useful in the management of diarrhea and other gastrointestinal diseases, and the use of these strains can be useful for prevention and treatment. Keywords:Lactobacillus, Dairyproducts, SalmonellaTyphimurium, E.coli, Helicobacter pylori

The Frequency of Streptococcus Mutans and Lactobacillus spp.in 3-5-year- old Children with and without Dental Caries

Z Moulana

Abstract Background and Objective: The high occurrence of early childhood caries (ECC) is one of the most common problems in children dentistry. Streptococcus mutans and lactobacilli.spp are closely associated with the development of early childhood caries (ECC). Therefore, the aim of this study was to investigate the frequency of streptococcus mutans and lactobacilli.spp in 3-5 –year- old children with and without dental caries. Material and Methods: This case-control study was conducted on 60 children aged 3 -5-years, without any history of systemic disease, who had not received any antibiotic therapy and fluoride usage during the last month. The cases were divided into three groups of early childhood caries, usual caries and caries-free. The infected dentin samples were collected from cervical and proximal in ECC and usual caries group, respectively. Also in all the three groups, the samples of dental plaque from buccal surfaces were collected and immediately immersed into Brain Heart Infusion (BHI) broth medium. After that, the diluted sample was plated onto MitisSalivarius agar (Difco) for detecting streptococcus mutans and Rogosa agar (Difco) for detecting lactobacilli.spp. Data were analyzed by Chi- Square and ANOVA. Results: of the samples taken from dental plaque, S. mutans is observed in 90% of ECC, 80% of proxymolcarries and in 25% of caries-free individuals. Based on the results, the presence of S.mutans in the group of caries is significantly higher than that of without caries, but there is no significant difference between the two groups of caries. In addition, lactobacill.spp is isolated from 60% ofthe samples, but there is no significant difference between two groups of caries and caries-free. The average of decayed, missing and filled teeth (DMFT) index in the group with early childhood caries (6.95±1.572) and usual caries (5.80±1.105) is different significantly. Pearson correlation test shows that there is a significant relationship between caries experience and the amount of S.mutans, but there isn’t this relationship withlactobacill.spp. Conclusion: The findings show that the children with early decay have the higher level of S.mutans in their dental plaque. The most common strain isolated from dentin, without consideration of their location, is S.mutans. Key words: Early dental care, Dental caries, DMFT index, Streptococcus mutans, Lactobacilli.spp.

The Prevalenceof Antibiotic Resistance Pattern of Staphylococcus Aureus Isolated from Nasal Carriage of Surgical Ward’s Staff in ShahidrajaeeHospitalofTonekabon,Iran

A NasrollahiOmran

Abstract Background and Objective:Staphylococcusaureus is one of the important factors causing nosocomial infections. Typically 25-30 percent of healthy people carry the bacteria in their anterior nasal cavity. The physicians(50%), nurses (70%) and hospital staff (90%) are the carriers of this bacteria, leading to the infection of inpatients. The emergence of antibiotic-resistant Staphylococcus strains to vancomycin and methicillin has brought about several problems in treatment of the infections caused by Staphylococcus strains. Hence, we aimed to study the frequency of staphylococcus aureus carriers and resistance pattern among medical personnel of the surgical ward in ShahidRajaee hospital, Tonekabon. Material and Methods: this analytic-descriptive study was conducted on the samples taken from nasal carriage of medical staff of surgical ward (N=120). Antibiotic- resistant of Staphylococcus strains was assessed by antibiogram and disk diffusion (DAD), in accordance with CLSI standards. Results: of 34 (28.33%) who are nasal carriers of staphylococcus, 12 are over 30 years old and 24 under 30. Based on antibiogram, 1.97% of specimens are sensitive to Gentamicin and Co-trimoxazole, 1.94% to Ciprofloxacin, 2.88% to Vancomycin and 6.20% to Methicillin. In addition, 100% of specimens are resistant to Ampicillin, 1.97% to Penicillin and 2.88% to Amoxicillin. Four isolates areresistant,both to methicillin and vancomycin. Conclusion:In this study, the spectrum of S. aureus resistant and sensitive strains to some antibiotics is similar to other studies, but a dramatic increase is seen in the rate of MRSA and non-susceptible cases to vancomycin. The Effectiveness of Penicillin, Amoxicillin and Ampicillin is still very low on S. aureus samples. Key words: Prevalance Resistance Pattern, Staphylococcus aureus, Medical Staff, Nasal Cavity, Tonekabon

Strategic Planning to Establish Quality System in Medical Laboratory

S Royani

Abstract Background and Objective: Every organization requires ongoing evaluation of existing conditions. The purpose of this study is to assess and analyze the standards and criteria that each Laboratory system is required to observe and upgrade them, to determine the gap between the ideal and the current status and finally to present the strategy and executive plan in order to achieve the desirable status. Material and Methods: This study was performed in a medical diagnostic laboratory in Gorgan by using the quality system checklist related to medical diagnostic laboratories, which was revised in 2009. Internal evaluation matrix (Internal Factor Evaluation) was used to examine the main factors in the context of establishing a quality management system in a clinical laboratory.After examining the factors, determining the laboratory status, recording the results of monitoring (in terms of strengths and weaknesses) and determining the gap between existing and desirable status, we provided the appropriate and effective solutions in line with defined standard. Results: of 164 items thatshould have been done in the first assessment, 111 (67.7%) items are in performed group and 53 (32.3%) are not in. After compiling and running a plan, 147 (89.6%) are performed, 15 (9.2%) needed to be modified and 2 (1.2%) still not performed. It is evident that a significant difference (p< 0.05) and a tangible improvement in current problems are seen after establishing the qualitysystem in the laboratory. The lab equipment, lab space and facilities, pre-examination process, testing process and post-examination process are considered .Regarding the health and safety in the laboratory, lab equipment, lab space and facilities, pre-examination process, testing process and post-examination process, no significant difference is observed between before and after the implementation of the program. Conclusion:after establishing the quality system in laboratory, a significant difference and tangible improvement in the current problems are observed. It is implied the importance of pre-planned responses to problems and the performance of strategic planning. Keywords: Strategic Planning, Medical Diagnostic Laboratory, Quality Control

An Overview of theLaboratory Diagnostic Procedures of Visceral Leishmaniasis (Kala-Azar)

M Fakhar

Abstract Visceral leishmaniasis (Kala-azar) is a systemic infection disease that can be diagnosed by some invasive procedures such as splenic, liver biopsy or bone marrow aspiration, whichare determined as the gold standards for diagnosing of this disease. At present, a variety of noninvasive tests having different specificities and sensitivities are available for the diagnosis of visceral leishmaniasis. Direct agglutination test (DAT) can be an appropriate and applicable method provided that proper antigens are prepared. The rapid rK39 strip test (for detection of antigen) can be used for diagnosis of visceral leishmaniasis (VL), which is suitable for acute forms of disease in the field. Other tests, such as rapid KATEX strip test (for detection of antigen) and polymerase chain reaction (PCR), which are recently recommended for diagnosis and prognosis of visceral leishmaniasis, are the simple, inexpensive and easily available under field conditions.This review article focuses on different, novel and current procedures for the diagnosis of visceral leishmaniasis. Key words: Laboratory diagnosis,visceralleishmaniasis, Kala-azar,rk39, Katex, PCR

Comparison ofβhCG Serum level in mild pre-eclampsia and healthy pregnant women

A Tabandeh

Abstract Background and Objective: Pre-eclampsia is the most common cause of maternal and neonatal mortality. Recent studies show that there may be a relationship between chorionic gonadotropine (HCG) and pre-eclampsia. Thus we aimed at assessing the serum level of βhCG after 28th week of pregnancy in mild pre-eclamptic and healthy pregnant women. Material and Methods: In this case-control study, we measured the level of βhCG in 26 pre-eclamptic and 26 normal pregnant women. The difference between groups was analyzed by Mann-Whitney test (P≤ 0.05). Results: The mean of βhCG in the case (73192 ± 42956 IU/L) and control group (34038 ± 21272 IU/L) is different significantly (P= 0.001). Considering mother’s age, gestational age and parity, there is a significant difference between the two groups (P=0.001). Conclusion: The results of our study show that the higher level of βhCG, the greater the risk of pre-eclampsia. Key words: Pre eclampsia, βhCG, Parity

SeroepidemiologicalPrevalence of Helicobacter Pylori of the Patients Referred to the Central Laboratory in South of Tehran, 2010

Abstract Background and Objective: Helicobacterpylorus is the major cause ofinflammation andulcer instomach, and immunoglobulin IgG is one of the antibodies produced against it, which is important in the course and diagnosis of the previoussufferers. The awareness of the prevalence of this disease can be helpful for the physicians to choose the way of treatment. Material and Methods: In these cross-sectional study, 516patients referred to laboratory was studied. After separating the serum, Antibody Helicobacter pylori IgGtest wasdone by ELISA method. Results: of 516, 156 (30.2 %) of the patients have a positive result, 51(32.7%) are males and 105 (67.3 %) are females. Positive percent of males (43.5%) is greater than females (26.5%).Over-45-year-old women (9.8 %) have the highest percentage of disease titers. Conclusion: The percent of positive cases in men is more than that the women. Over-45-year-old women (9.8 %) have the highest percentage of positive case. Keywords: Helicobacter pylori IgG, Tehran, ELISA