Medical Laboratory science علوم المختبرات الطبية
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- ItemAntibiotic Resistance Patterns and Molecular Typing of Acinetobacter baumannii Clinical Isolates and Hospital Environment at Alia Governmental Hospital, Palestine(Al-Quds University, 2024-05-28) Donia Mohammad Mosbah Khderat; دنيا محمد مصباح خضيراتBackground:In recent years, Acinetobacter baumanniireceived increasing attention;due to theircabability of acquiring multi-resistance to antibiotics and their potential to cause disease in humans. It is also known for its intrinsic antimicrobial resistance mechanisms: several studies documentedthe increase in the percentage of resistance to antimicrobial agents in four majorantimicrobialclasses: Fluoroquinolones,aminoglycosides, β-lactams including Carbapenems. The increase in Carbapenems resistance is of important concern, as thisclass of antibiotics isusually considered the last choice in treating resistant A.baumannii.This opportunistic pathogen able to survive in the environment under unfavorable conditions for prolonged periods,this contributes to its increased virulance. Thus, the hospital environment may serve asa reservoir for the resistantA. baumannii strainsallowing easily spread in hospital,serve as a nosocomial infections in patients, healthcare personnel, and visitors. Aims: This study aimed at characterization of A. baumanniiisolates from inpatients and hospital environments.To find out their antimicrobial resistance pattern, detect Carbapenems resistant A.baumannii(CRAB), and to find out genetic relatedness between isolates using random amplification of polymorphic DNA (RAPD) typing technique. Methodology: In this study, a total of 24 clinical isolates obtained from various clinical specimenssuch as wounds, blood, urine, earswabs and sputumduring the period April1, 2023, and December 31, 2023fromAlia Governmental Hospital, Hebron,Palestine. The Vitek2 system was used for identification and antibiotic susceptibility testing. Concurrently a total of 126 specimen were collected from different environments of the hospital identified as A. baumanniiusing conventional identification methodsand CHROMagar Acinetobacterselective medium. The isolates then tested for antibiotic susceptibility testing using disk diffusion methods.All isolates (both clinical and v environmental) were tested for the presence ofblaOXA-51 gene using PCR. And finally, the genetic relatedness between isolates was determined using RAPDtyping technique. Results:Twenty fourclinical isolates and 29 environmental isolates were identified as A. baumannii. All isolates were confirmed to be A. baumanniiby detecting blaOXA-51-like gene.Among the environmental isolates patient’s beds were the most contaminated site (20.6%, 6/29),while the largest number of clinical isolates were obtained from wounds (10 isolates: 41.7%). The majority of clinical and environmental isolates were collected from intensive care unit (ICU).Multidrug resistant A.baumannii (MDRAB) was detected in approximately 74% of clinical isolates and in 100% of environmental isolates.Co-trimoxazole (COT) was the most effective antibiotic against both clinical and environmentalA.baumannii isolates. For other antibiotics, clinical and environmentalA.baumanniiisolates showed high resistance rates:86.2% and 96.6% of environmentalisolates were resistant to imipenem (IM) and meropenem (MEM) respectively,while clinical isolates had a resistant rate of 72.7% and 77.3% to IM and MEM, respectively.Using the RAPD-PCR typing technique, four clusters grouped A-D had more than one isolateand the remaining isolates each had unique pattern. Some RAPD clusters were seen with environmental as well as clinical isolates. Conclusion and recomendations:Thisstudy clearly suggests that the rate of MDRAB and CRAB are increasing and considered an important cause of nosocomial infections.The relatednessbetween clinical and environmental isolates and the detection of high rate of antimicrobial-resistant A.baumanniiin various hospital environments suggested the potential role of the hospital environments through various clinical activities in the cross contamination of A.baumanniiinfections, particularly in the ICU. These findings highlight the importance of identifying A.baumannii infections early and implementing very strict infection control strategies.It is recommended that hospital wards must be vigorously sanitized and disinfected periodically.
- ItemInvestigating β-Thalassemia Major Cases Emerged After Obligatory Premarital Testing in Palestine(Al-Quds University, 2023-11-11) Mais Khader Yousef Al Shatleh; ميس خضر يوسف الشتلةβ-Thalassemia major is a prevalent autosomal recessive disorder worldwide, affecting approximately 3-4% of the Palestinian population. Management of this condition poses significant challenges, particularly in resource-constrained settings. This study aims to investigate the factors contributing to the emergence of new cases of β-thalassemia major after the implementation of obligatory premarital screening, thereby informing strategies for prevention and intervention. Methods We conducted a cross-sectional case-series study that included all patients diagnosed with β-thalassemia born in or after 2010 in the West Bank. 69 eligible patients from 62 families were included in the study. The data used in this study were collected through a comprehensive questionnaire covering the demographic and medical information of each thalassemic child born after 2010 in the family, familial sociodemographic characteristics, background characteristics of the parents (fathers and mothers), premarital screening, and the knowledge, attitudes, and practices (KAP) of parents (mothers and fathers). In addition, we collected blood samples from 68 children and 62 parents for hematological assessment (complete blood count and hemoglobin electrophoresis). Results The largest proportion of emerging thalassemia cases were from the northern region and resided in rural localities. 71% of the cases were from families married before 2010. 56.5% of the parents reported undergoing premarital screening tests. The proportion of parents who underwent premarital screening differed significantly by type of locality, year of marriage, and age at marriage. In addition, investigating the self-reported results of the premarital screening tests of each couple, we have found that among 24 partners who did not get tested, 22 were married before 2010 and 19 had children with β-thalassemia major. Furthermore, among 12 couples who reported that the two partners were tested and were non-carriers, 4 couples had children with β-thalassemia major, 4 had children with sickle cell thalassemia, and 4 had children with thalassemia intermedia. Overall, the proportion of children with thalassemia major was lower by 20% among parents who married in/after 2010. On the other hand, comparison between the self-reported results of premarital screening and the results of the hematological assessment (mean capsular volume), a total of 24 out of 32 parents had discrepancies in their results. the hematological assessment also showed that 3 out of 62 parents had normal MCV, all of which had high HbS and were parents of children with sickle cell thalassemia. Also, 8 parents had both low MCV and high HbS and one parent had low MCV and high HbC and was a parent of two children with hemoglobin SC disease. Assessment of knowledge showed that all parents had adequate knowledge about thalassemia, 51.7% of the parents had poor overall attitudes, and 76.3% had poor attitude scores towards the termination of pregnancy. Furthermore, 47.4% of the parents had good overall practice scores. Knowledge scores were positively correlated with attitude scores (r=300; p-value=0.01) but not with practice scores (r=0.058; p-value=0.543). Attitude score towards prenatal diagnosis and overall practice scores were also positively correlated (r=0.271; p-value=0.003). Conclusions This study highlights the pressing need for proactive measures to address the prevalence of hemoglobinopathies in Palestine. Findings highlight the crucial role of Health Education and Awareness Programs, aimed at disseminating information about thalassemia, its inheritance patterns, and preventive measures. Additionally, the screening criteria should be revised to include screening for other hemoglobinopathies, and the screening must be performed in Designated Receptions Centers where trained staff perform and interpret the results in accordance with standard laboratory diagnostic protocols. Strengthening pre-marital counseling and screening services is imperative to ensure comprehensive coverage. Early detection, accurate carrier identification, and informed decision-making regarding marriage and family planning should be central in preventive efforts. Continuous training for healthcare professionals and community workers is crucial to effectively prevent and manage thalassemia.
- ItemThe Prevalence, Diagnosis, and Antifungal Susceptibility of Candida Species in Palestine(Al-Quds University, 2023-08-20) Hanaa Khairaldeen Mohammed Baniodeh; هناء خيرالدين محمد بنيعودةBackground: Candida spp are the single most common cause of opportunistic fungal infection.Various healthcare implications are associated with Candida infections, including morbidity and mortality. The significantly increased occurrence of Candida species as human pathogens can be attributed to improved identification techniques. In Palestine, minimal data have been reported about Candida infection. Methods: We performed our study at two hospitals in Palestine ( Istishari Arab Hospital, and Najah National University Hospital ). All patients diagnosed with candidiasis during the year 2022 have participated in the study. The prevalence of Candida spp,their distribution and the activity of selected antifungals against Candida pathogens were statistically analyzed using Microsoft Excel Version (16.74). In combination with phenotypic properties, Candida isolates were identified and tested for antifungal susceptibility using the colorimetric VITEK-2 Compact system at two hospitals. Results : Our results showed that the prevalence of Candida spp among infected samples was 11.6%. Twelve different Candida spp were identified. Among these isolates, C. albicans (49.88%) was the most frequent, followed by C. glabrata (17.3%), C. tropicalis (14.83%), C. parapsilosis (5.16%), C. krusei (3.82%), C.auris(2.69%) ,C.dubliniensis (2.24%), C.ciferrii (1.79%) C.lusitaniae (0.89%), C. guilliermondii (0.67%), C.Kefyer (0.44%) and C.spherica(0.22%). Among C. albicans, all isolates were 100% susceptible to fluconazole, and micafungin .The susceptibility rates to Amphotericine B and flucytosine were 95 % and 99 % respectively. As a comparison, the susceptibility rates of non-albicans candida to fluconazole, voriconazole, amphotericine B, caspofungin ,flucytosine and micafungin were 70 %, 99 %, 97 %, ,72% , 92% and 100 %, respectively. Patients in the intensive care unit are more susceptible to candida infections than patients in other hospital departments. Conclusions: Four pathogens are responsible for the most invasive infections: Candida albicans, Candida glabrata, Candida tropicalis, and Candida parapsilosis. Those in an immunocompromised state are more likely to develop Candida infections. A notable characteristic of this study was the high frequency of non-albicans Candida Spp, which were often more resistant to antifungal agents. A quick and accurate system like Vitek 2 compact was suggested for careful species identification of clinical isolates of Candida. We suggest that continued surveillance of species distribution and susceptibility to antifungals will enhance future burden estimates and assist in evaluating preventative measures' effectiveness.
- ItemHuman Parvovirus B19 Epidemiology, Genotyping, Patients’ Hematological Presentation and Clinical Manifestations in Children in Southern Palestine(Al-Quds University, 2023-09-03) Miral Munther Badawi Abdo; ميرال منذر بدوي عبدهBackground: Human parvovirus (B19V) is a virus that is usually asymptomatic in healthy patients but can sometimes be symptomatic, especially in patients who have a previous underlying disease such as hematological and cardiopulmonary diseases. B19V targets the erythroid progenitor cells by attaching to the P antigen causing slapped cheek syndrome and may cause transient aplastic anemia. The aim of the study was to check the prevalence of B19V, the most common genotypes, its effect on CBC and the clinical symptoms especially transient aplastic crisis for the first time in Palestine. Methods: B19V was studied in children retrospectively from February 2014 to December 2022 in Caritas baby hospital in Bethlehem. Real-Time PCR for B19V was done on blood EDTA tube for 905 suspected patient samples out of which 28 patients tested positive for B19V (3%). The 28 B19V cases were compared with 32 B19V negative controls. Both groups were checked for social demographics, clinical symptoms, hematological parameters (CBC), and other lab tests (CRP, AST, and ALT). Moreover, genotyping was attempted on all RT-PCR positive B19V cases. This was done by amplifying B19V VP2 gene using VP2 special primers followed by Sanger sequencing. Results: As for social demographics, being a female or a male does not affect the chance of being infected with B19V, in addition to the age of the patient and the place of inhabitant. Regarding clinical manifestations some variables showed significant statistical association with B19V infection such as respiratory problems (p =0.046), malaise (p =0.005), and neurological issues (p =0.043); however, having fever, rash and anemia did not have any statistically significant association with B19V although 2 patients had severe anemia: one patient was diagnosed with pure red cell aplasia and another patient was diagnosed aplastic anemia. A statistically significant association was also found in certain CBC parameters, including WBC (p =0.003), platelets (p =0.018) and Hb (p =0.019, OD=9.6, 95% confidence interval: 1.45,63.5), notably patients with Hb < 10 mg/dl are 9.6 times more likely to test positive for B19V compared to Hb>10. CRP, AST, and ALT had no significant association with B19V, although all the B19V positive patients had CRP above 6 mg/dl. Of the patients analyzed, 40.7% of the cases had previous underlying disease which was significantly associated with B19V infection (p =0.003, OD=10.3, 95% confidence interval: 2 ,52.3), thus individuals with a previous underlying disease are 10.3 times more likely to test positive for the B19V compared to previously healthy patients. Moreover, 37% of the cases had coinfections of B19V and another microorganism. As for genotyping, of the 28 positive samples analyzed VP2 fragment was amplified from 5 samples (17.8%) that had high B19V viral load and the phylogenetic tree revealed that the most common genotype of B19V in Palestine is genotype 1.Conclusion: To conclude, this retrospective case-control study is the first study to check B19V in Palestine which provided valuable insights and more understanding about the virus in the country.
- ItemEvaluation of Western blot analysis for the detection of COVID-19 infected and vaccinated serum samples against different viral specific antigen preparations(Al-Quds University, 2023-01-15) Aseel Basem Sami Eqneiby; أسيل باسم سامي اقنيبيمقدمة:فيروس كورونا هوعبارة عن فيروس يصيب الجهاز التنفسي ويطلق عليه اسم المتلازمة التنفسية الحادة (SARS-Cov-2) , ينكون هذا الفايروس من جينوم RNA أحادي موجب الاتجاه (+ ssRNA) حيث يترواح حجمه بين ال 30 و ال 32 كيلو بايت تتضمن مجموعة من البروتينات السطحية والوظيفية , يقوم فيروس كورونا ب احداث المرض والوصول الى خلايا الجسم من خلال ارتباطه بالخلايا السطحية المعروفة ب اسم ACE2 والموجودة في خلايا الرئة , الهدف الرئيسي من هذه الدراسة هو استخدام الانتيجينات المختلفة لفيروس كورونا و التي تم الحصول عليها مخبريا لفحص وجود لاجسام المضادة داخل مجموعة من العينات الماخوذة من مجموعات مختلفة من المرضى ب استخدام تقنية ال Western-blot الطريقة: تم استخدام مجموعة من ال plaques تسمى ال M13 phase plaquesوالتي تحتوي على 12 منطقة من الاحماض الامينية ضد مجموعة من الامصال الإيجابية المجمعة ل مرضى COVID-19, تم استخدام مجموعتان من ال 12 amino acids والبروتينات المصنعة في تحليل ال Western-blot, كانت الاصال المستخدمة في هذه الدراسة ثلاثة أنواع : (20 عينة مصل COVID-19 موجبة ومطعمة ، 10 عينات مصل إيجابية غير ملقحة ، وبعض عينات المصل السلبية). استنتاج النتائج: بشكل عام استخدام phages M13 التي تحتوي على 12 حمضًا أمينيًا تم اختيارها باستخدام الأمصال المصابة بفيروس COVID-19 ؛ ثبت أنه أكثر تفاعلًا مع عينات مرضى ال COVID-19 المحصنة وحتى المصابة وغير الملقحة ، وهذا مقارنة باستخدام مستضدات البروتينات المؤتلف السنبلة ال s protein أو الغشائية ال m protein . وقد لوحظ بوضوح أن هذه الانتيجينات المختلفة لم تتفاعل سوى مع IgG وليس IgM ، مما يشير إلى فائدتها في الكشف عن الحالات التي تحتوي على عيارات عالية من الأجسام المضادة IgG وليس في حالات عدوى COVID-19 الحادة. يمكن أن تؤكد هذه النتيجة على إمكانية استخدام phages M13 التي تحتوي على 12 من الأحماض الأمينية في تقييم إجراءات التطعيم ضد COVID-19.