Evaluation of the discriminatory power of spoligotyping and 19-locus mycobacterial interspersed repetitive unit-variable number of tandem repeat analysis (MIRU-VNTR) of Mycobacterium bovis strains isolated from cattle in Algeria

Bovine tuberculosis (bTB) caused by Mycobacterium (M.) bovis and M. caprae is a transmissible disease of livestock, notifiable to the World Organization for Animal Health (OIE). BTB particularly affects cattle and small ruminants and can be transmitted to humans thereby posing a significant threat to veterinary and public health worldwide. M. bovis is the principal cause of bTB in Algeria. In order to better understand the route of spreading and elaborate an eradication program, isolation and characterization of mycobacteria from Algerian cattle was performed. Sixty strains belonging to the M. tuberculosis complex were analyzed by spoligotyping, thereof 42 by 19-locus-MIRU-VNTR-typing. Spoligotyping revealed 16 distinguishable patterns (Hunter-Gaston discriminatory index [HGDI] of 0.8294), with types SB0120 (n = 20) and SB0121 (n = 13) being the most frequent patterns, representing 55% of the strains. Analyses based on 19-locus-MIRU-VNTR yielded 32 different profiles, five clusters and one orphan pattern, showing higher discriminatory power (HGDI = 0.9779) than spoligotyping. Seven VNTR-loci [VNTR 577 (alias ETR C), 2163b (QU11b), 2165 (ETR A), 2461 (ETR B), 3007 (MIRU 27), 2163a (QUB11a) and 3232 (QUB 3232)] were the most discriminative loci (HGDI ˃ 0.50). In conclusion, 19-locus-MIRU-VNTR yielded more information than spoligotyping concerning molecular differentiation of strains and better supports the elucidation of transmission routes of M. bovis between Algerian cattle herds.

SOAL KEMAMPUAN PEMECAHAN MASALAH MATEMATIS BERKONTEKS KEARIFAN LOKAL BENGKULU

This study aims to produce products in the form of mathematical problem-solving ability questions based on Bengkulu local wisdom that are valid, clear, and have a good level of difficulty and discriminatory index. The subjects in this study were 30 grade VIII students of SMPN 07 Bengkulu Selatan. This development research was conducted using the Tessmer model which consisted of preliminary, self-evaluation, expert review, one-to-one, and small group stages. The data analysis used was qualitative analysis at the expert review stage to determine the validation of the questions, both in terms of content, construction, and language; one-to-one process analysis to find out the clarity of the questions, and quantitative analysis of the results of the small group to determine the characteristics of good questions based on the level of difficulty and the index of discriminating power of the questions. Based on the results of data analysis, this study produced 7 standardized questions from 10 questions that had gone through the expert review, one-to-one, and small group stages. Standardization can be seen from the level of difficulty and a good item discriminatory index.

Desain soal-soal kemampuan literasi matematis untuk siswa SMP

This research aimed to produce questions of mathematical literacy ability of Mathematical literacy ability on Triangular material for Class VII Junior High School students that are valid and practical and have a good level of difficulty and discriminatory index. This research used the Tessmer development model. Tessmer's development stages include preliminary, self-evaluation, expert review, one-to-one, and small group. The test subjects of this study were class VII students of SMP Negeri 02 Kepahiang, totaling 30 students. The selection of test subjects includes high, medium and low abilities. The data analysis technique of this study used qualitative analysis at the expert review and one to one stage and quantitative analysis at the small group stage by analyzing items based on the level of difficulty and discriminating power index. The results of the research show that the 10 questions developed, 10 questions with indicators of mathematical literacy ability on triangular material are valid and practical, 7 questions are said to be good based on the level of difficulty and discriminatory index, so that they can be used by all students of SMPN 02 Kepahiang.

Pre and Post examination evaluation of MCQs and SEQs Items in Undergraduate MBBS mock examination.

Objectives: To evaluate the quality of MCQs and SEQs pre and post Mock examination of physiology and biochemistry and correlation between the scores of both at private medical college Lahore. Study Design: Descriptive study and convenient sampling was done. Setting: Fatima Memorial College of Medicine and Dentistry, Lahore. Period: September 2016 to September 2017. Material & Methods: 149 students in Physiology and 143 in biochemistry took Mock examination. 45 MCQs and 9 SEQs each in biochemistry and physiology were prepared according to the table of specification provided by University of Health Sciences (UHS) Lahore, Pakistan. Item assessment according to Blooms taxonomy was done and item flaws identified with cover test done for structural validity of the paper before the exam by two medical educationists. Item analysis with difficulty index, discrimination index and distraction efficiency were done post examination. Results: 84.4% in physiology and 51.1% in biochemistry were of recall type (C1 level according to Blooms taxonomy), and 58% SEQs in physiology and 50% in biochemistry were C1. 20% and 28% MCQs in physiology and biochemistry respectively pass cover test and were without item writing flaws. Difficulty index shows that 53.3% in physiology and 48.8% in biochemistry needs modifications. 48.8% of MCQs in physiology and 15.5% in biochemistry need modification in discriminatory index. Similarly 59.5% and 64.0 % of MCQs had functional distractors in physiology and biochemistry respectively. Conclusion: The study concluded that Mock examination in subjects of physiology and biochemistry had item writing flaws, more MCQs were of C1 level and showed majority items in good difficulty, discriminatory index with two third functional distractors were present

Genotypes of Mycobacterium tuberculosis isolates circulating in Shaanxi Province, China

Objectives The prevalence of drug-resistant TB in Shaanxi Province is higher than other areas. This study was aimed to investigate the genetic diversity and epidemiology of Mycobacterium tuberculosis clinical strains in Shaanxi Province, China. Methods From January to December 2016, a total of 298 Mycobacterium tuberculosis clinical isolates from smear-positive pulmonary tuberculosis patients were genotyped by Mcspoligotyping and 15-locus VNTR. Results We found that the Beijing family strains was the most prominent family(81.54%, 243/298). Other family strains included T family(9.06%, 27/298), U family(0.67%, 2/298), LAM9 family(0.34%, 1/298) and Manu family(0.34%, 1/298). The rates of multidrug-resistant (MDR) M.Tuberculosis, age, type of case and education between Beijing and non-Beijing family strains were not statistically different, while the distribution in the three different regions among these was statistically significant. VNTR results showed that strains were classified into 280 genotypes, and 33 (11.07%) strains could be grouped into 14 clusters. 11 of the 15-VNTR loci were highly or moderately discriminative according to the Hunter-Gaston discriminatory index. Conclusions We concluded that the Beijing family genotype was the most prevalent genotype and 15-locus VNTR typing might be suitable for genotyping of M. tuberculosis in Shaanxi Province. There was less association between Beijing family genotypes and drug resistance in our study area.

Typing of ΦSP–3 lytic Salmonella bacteriophages obtained from various fecal sources

Although several reports are available on both ΦSP–1 and ΦSP–3 lytic Salmonella bacteriophages obtained from poultry, further research is required to study the effectiveness of ΦSP–3 type on serovars isolated from other sources. In the present study, we aimed to isolate bacteriophages from 8 serovars previously obtained from 869 fecal samples (calf, dairy cow, buffalo, and camel), genotype the bacteriophages, and detect the cross-lytic activities of the bacteriophages on Salmonella enterica subsp. enterica serovar Kentucky, S.Anatum, and S.Muenchen. A total of 16 bacteriophages were detected as ΦSP–3 type via PCR. The Hunter-Gaston Discriminatory Index of SDS-PAGE was calculated to be 0.825. Determination of multiplicity of infection (MOI) values were different for each bacteriophage according to the cross-lytic activity assessment. The MOI of the most effective S. Kentucky bacteriophage was 79.11 μg/mL for 2.5×104 cells, whereas that of the most ineffective S.Muenchen bacteriophage was 1.142 μg/μL for 2.5×104 cells. In conclusion, it was assumed that owing to the high and cross-lytic activity of the S. Kentucky bacteriophage, it has a larger host range, which differs in the lytic activities of each bacteriophage, despite being the same serovar, and that calf feces is the most important source for obtaining Salmonella bacteriophages.

Comparative Evaluation of Band-Based Genotyping Methods for Mycobacterium intracellulare and Its Application for Epidemiological Analysis

Mycobacterium intracellulare is a leading cause of nontuberculous mycobacterial pulmonary disease, with a rapidly increasing prevalence worldwide. This bacterium, commonly distributed in soil and water, is known to be transmitted through the environment rather than between people. Therefore, it is imperative to establish distinguishable genotyping methods to understand the clinical outcome, disease relapses, and epidemiology. Therefore, in this study, representative band-based genotyping methods were performed using M. intracellualre clinical isolates, and their Hunter–Gaston discriminatory index (HGDI) was 0.947, 0.994, and 1 for variable number tandem repetition (VNTR), VNTR-mycobacterial interspersed repetitive units, pulsed field gel electrophoresis, and repetitive sequence based-PCR, respectively. Although VNTR showed relatively low HGDI, co-infection with other M. intracellualre strains could be determined by loci showing allele diversity from 0 to 0.69. Additionally, genetic distance of clinical isolates from Gyeongnam/Korea, and other regions/countries were visualized by minimum spanning tree (MST) using the globally available VNTR profiles. The results of MST revealed that M. intracellulare isolated from patients in Gyeongnam/Korea had specific VNTR genotypes, which may be evidence of the geographic distribution of M. intracellulare specific genotypes. The comparative results of genotyping techniques and geographical characteristics in this study may provide fundamental information for the epidemiology of M. intracellulare.

233. The Epidemiology, Genomics, and Evolution of Staphylococcus aureus in Northeast Ohio

Background Infections due to S. aureus result in significant morbidity, mortality, and healthcare expense. We sought to identify the strains of S. aureus causing infections in hospitalized patients in Northeast Ohio and determine whether they are reflective of the S. aureus strains present in the surrounding environment. Methods The study was approved by the Institutional Review Board at Cleveland Clinic Akron General. Clinical S. aureus isolates (n = 300) were cultured and PCR was used to amplify the staphylococcus protein A (spa), Panton–Valentine Leukocidin (PVL), and mecA genes. The clinical spa types were compared with ones from our data base of S. aureus strains previously collected and sequenced from the community and environment in Northeast Ohio. Results A total of 51 spa types were detected from 129 S. aureus clinical isolates (discriminatory index, 0.876; 95% confidence interval [CI], 0.827–0.925; Table 1). The most common spa types were t008 (42/129, 32.6%), t002 (16/129, 12.4%), and t334 (6/129, 4.7%). In comparison, the most frequently detected spa types from the environmental samples were t189 (40/257, 15.6%), t002 (16/257, 6.2%), and t008 (11/257, 4.3%). Among the S. aureus isolates (n = 146), 45 were PVL-positive (30.8%) and 94 (66.7%) carried mecA. Of the 42 t008 (ST8/USA300; a common community-associated strain) isolates, 35 (83.3%) were methicillin-resistant S. aureus (MRSA) (based on the presence of the mecA gene) and 25 (59.5%) were PVL-positive. Thirteen of the sixteen (81.2%) t002 (ST5/USA100; a common hospital-associated strain) were MRSA and only one (6.2%) was PVL-positive. Conclusion There is considerable overlap of S. aureus strains present in clinical samples with those found in the environment. This finding should draw attention to the need for more effective prevention strategies to reduce the risk of transmission of S. aureus, including MRSA, in the environment to humans. Disclosures All authors: No reported disclosures.

Impact of distractors in item analysis of multiple choice questions

Background: Item analysis is a quality assurance of examining the performance of the individual test items that measures the validity and reliability of exams. This study was performed to evaluate the quality of the test items with respect to their performance on difficulty index (DFI), Discriminatory index (DI) and assessment of functional and non-functional distractors (FD and NFD).Methods: This study was performed on the summative examination undertaken by 113 students. The analyses include 120 one best answers (OBAs) and 360 distractors.Results: Out of the 360 distractors, 85 distractors were chosen by less than 5% with the distractor efficiency of 23.6%. About 47 (13%) items had no NFDs while 51 (14%), 30 (8.3%), and 4 (1.1%) items contained 1, 2, and 3 NFDs respectively. Majority of the items showed excellent difficulty index (50.4%, n=42) and fair discrimination (37%, n=33). The questions with excellent difficulty index and discriminatory index showed statistical significance with 1NFD and 2 NFD (p=0.03).Conclusions: The post evaluation of item performance in any exam in one of the quality assurance method of identifying the best performing item for quality question bank. The distractor efficiency gives information on the overall quality of item.

Evaluation of BOX-PCR and ERIC-PCR as Molecular Typing Tools for PathogenicLeptospira

In the last decades, leptospirosis had gained public health concern due to morbidity and mortality rates caused by pathogenicLeptospira. The need for rapid and robust molecular typing methods to differentiate this zoonotic pathogen is of utmost importance. Various studies had been conducted to determine the genetic relatedness ofLeptospiraisolates using molecular typing methods. In this study, 29 pathogenicLeptospiraisolates from rat, soil, and water samples in Sarawak, Malaysia, were characterized using BOX-PCR and ERIC-PCR. The effectiveness of these two methods with regard to the ease of interpretation, reproducibility, typeability, and discriminatory power was also being evaluated. Using BOX-PCR, six clusters and 3 single isolates were defined at a genetic distance percentage of 11.2%. ERIC-PCR clustered the isolates into 6 clusters and 2 single isolates at a genetic distance percentage of 6.8%. Both BOX-PCR and ERIC-PCR produced comparable results though the discriminatory index for ERIC-PCR (0.826) was higher than that for BOX-PCR (0.809). From the constructed dendrogram, it could be summarized that the isolates in this study were highly heterogeneous and genetically diverse. The findings from this study indicated that there is no genetic relatedness among the pathogenicLeptospiraisolates in relation to the locality, source, and identity, with some exceptions. Out of the 29 pathogenicLeptospiraisolates studied, BOX-PCR and ERIC-PCR successfully discriminated 4 isolates (2 isolates each) into the same cluster in relation to sample sources, as well as 2 isolates into the same cluster in association with the sample locality. Future studies shall incorporate the use of other molecular typing methods to make a more thorough comparison on the genetic relatedness of pathogenicLeptospira.