Evaluation of PCR pncA-restriction fragment length polymorphism and PCR amplification of genomic regions of difference for the identification of M. bovis strains in lymph nodes cultures

Background: A rapid accurate identification of Mycobacterium bovis is essential for surveillance purposes. Objectives: A PCR pncA-Restriction Fragment Length Polymorphism (RFLP) and a multiplex PCR based on the detection of 3 regions of difference (RD-PCR): RD9, RD4 and RD1 were evaluated for the identification of M. bovis in lymph nodes cultures, in Tunisia, during 2013-2015. Methods: Eighty-two M. tuberculosis complex strains were identified using the biochemical tests, GenoType MTBC assay, PCR pncA-RFLP and RD-PCR. Results: The PCR pncA-RFLP showed that 54 M. bovis strains, identified by GenoType MTBC, had a mutation at position 169 of pncAi> gene. Twenty-eight strains did not show any mutation at this position 27 M. tuberculosis isolates and one M. caprae . The PCR pncA-RFLP had a sensitivity of 100.0% (95%CI: 93.3 -100.0) and a specificity of 100.0% (95%CI: 87.9-100.0) for identifying M. bovis. The RD-PCR showed that all M. bovis strains had the RD9 and RD4 deleted but presented RD1. RD-PCR also presented high sensitivity and specificity in detecting M. bovis strains (100.0%). Conclusions: PCR pncA-RFLP and RD-PCR represent very accurate and rapid tools to identify M. bovis. They can be easily implemented in each laboratory due to their low cost and easy use

Individual or Group-based Approach to the Assessment of Preschool Children: A Comparison using the INTERGROWTH-21st Neurodevelopment Assessment (INTER-NDA)

Introduction: It is unclear if the assessment of early child development can be carried out using a group approach, as opposed to individually.&#x0D; Objective: To compare scores obtained from children aged 22 to 26 months assessed either in small groups or individually using the INTERGROWTH-21st Neurodevelopment Assessment  (INTER-NDA), which measures cognition, language, motor skills, behavior, attention and socio-emotional reactivity.&#x0D; Methods: A small group based strategy for administering and scoring the INTER-NDA was developed. Thirty-six preschool children attending four Centros de Cuidado y Atención Infantil of the Sistema Nacional para el Desarrollo Integral de la Familia (DIF) of Mexico were assessed in small groups of three children by a teacher specifically trained in the INTER-NDA. A second teacher, unaware of the group results, assessed the children individually on a different day. The sex, age, weight, length and head circumference of the children at the time of assessment were recorded.&#x0D; Results: INTER-NDA domain scores for group and individual assessments were statistically significantly correlated (range r=0.35 to r=1.00) for all domains except receptive language (r=0.25, p=0.14). Bland-Altman analysis showed agreement between group and individual scores for the language, behavior, attention and socio-emotional reactivity domains, and consistency (but not agreement) between group and individual scores for the cognitive and motor domains. None of the differences between group and individual scores examined were statistically significant, even after adjusting for the children’s age, sex, nutritional status and location of the preschool.&#x0D;  &#x0D; Conclusion: INTER-NDA domain specific scores obtained following group and individual assessment of children aged 22 to 26 months are consistent. It is feasible for trained preschool teachers to administer INTER-NDA at both group and individual level.</jats:p

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

The Impact of Innovation on Store Managers’ Job Satisfaction: Evidence from Tunisian Retailing

International audienc

Comparison of pp65 antigenemia, quantitative PCR and DNA hybrid capture for detection of cytomegalovirus in transplant recipients and AIDS patients

International audienceThe cytomegalovirus (CMV) antigenemia assay has been used frequently for rapid diagnosis of CMV infection, and antigenemia threshold values are recommended for triggering preemptive therapy. Hybrid capture of CMV's DNA and quantitative polymerase chain reaction (qPCR) are increasingly being adopted for early detection of CMV. The performance of the antigenemia assay, qPCR in plasma and hybrid capture in leukocytes were compared in 110 immunocompromised patients (38 bone-marrow transplants, 50 renal transplants and 22 AIDS patients). The most sensitive test was hybrid capture for transplants, while antigenemia and the qPCR showed similar performance for patients with AIDS. QPCR and hybrid capture thresholds requiring antiviral therapy were calculated using a receiver-operating-characteristic curve for antigenemia values corresponding to 2 positive cells for bone-marrow transplants and to 10 positive cells for renal transplants and AIDS patients. These threshold values varied with the group of patients considered, with corresponding sensitivities higher than 86% and specificities higher than 76% for hybrid capture, and sensitivities higher than 61% and specificities higher than 75% for qPCR in plasma. Hybrid capture in leukocytes can substitute for antigenemia in the case of transplants, and qPCR in plasma can substitute for it in the case of AIDS patients. (C) 2007 Elsevier B.V. All rights reserved

Evaluation of GeneXpert MTB/RIF for the detection of Mycobacterium tuberculosis and resistance to rifampin in extra-pulmonary specimens

Aims and objectives: This study evaluated the performance of GeneXpert (GX) for direct detection of MTB in extra-pulmonary specimens and assessed the ability of the assay to detect resistance to RIF. Methods: 104 clinical samples from a tertiary hospital in Sfax, Tunisia were analyzed. Specimens were processed using GX, Ziehl Neelsen and auramine smear, conventional culture on LJ and MGIT >60 media, and drug phenotypic susceptibility testing on LJ and MGIT 960. The diagnosis was made based on clinical, radiological, microbiological, pathological and therapeutic criteria. Results: In total, 51 patients were considered tuberculous. The PCR result obtained in less than two hours was positive for 46 samples and rifampicin was sensitive to all these cases. The sensitivity, specificity, positive predictive value and negative predictive value of GX were 90%, 60.6%, 19.6% and 98.3% compared with direct examination, 84.8%, 74.6%, 60.9% and 91.4% compared with culture and 84.3%, 94.3%, 93.4%, 86.2% compared with final diagnosis. The sensitivity of the GXt was 86% in biopsies and 75% in pus, collections and in biological fluids. Conclusion: The GX is a simple, rapid technique for real-time PCR and has increased the sensitivity of detection of Mycobacterium tuberculosis complex. It must be part of the diagnostic arsenal of tuberculosis without replacing conventional microbiological tools and allow early diagnosis and appropriate treatment

Diagnosis of lymph node tuberculosis using the GeneXpert MTB/RIF in Tunisia

Introduction: GeneXpert MTB/RIF is a fully-automated diagnostic molecular test which simultaneously detects tuberculosis (TB) and rifampicin (RIF) drug resistance. The purpose of this study is to evaluate the performance of the GeneXpert MTB/RIF test for the detection of Mycobacterium tuberculosis complex (MTBC) in lymph node specimens and to show the place of Mycobacterium bovis as a major cause of TB lymphadenitis. Material and methods: This study was conducted simultaneously in the National Reference Laboratory for Mycobacteria of Ariana and the Central Laboratory of Sfax, from January to December 2013. In total, 174 lymph node specimens were processed simultaneously for Ziehl–Neelsen, auramine and immuno-histochemical staining. Conventional culture on both Lowenstein–Jensen and liquid medium (Bactec MGIT 960 BD system) and the new molecular-based GeneXpert MTB/RIF assay system were performed. Positive cultures were confirmed using molecular identification (Genotype MTBC Hain Lifescience). Results: Among the 174 samples tested, the GeneXpert detected the DNA of MTBC in 134 samples (77%). Standard bacteriological assays, including AFB microscopy and culture, were positive, respectively, in 41 (23.6%) and 79 (45.4%) specimens. M. bovis was isolated in 76% of positive cultures. GeneXpert sensitivity and specificity results were assessed according to smear and culture results, clinical and histological findings. The sensitivity and specificity of the Xpert assay were 87.5% (126/144) and 73.3%, respectively. Conclusion: The implementation of the GeneXpert MTB/RIF assay may dramatically improve the rapid diagnosis of lymph node TB

Population Diversity of Staphylococcus intermedius Isolates from Various Host Species: Typing by 16S-23S Intergenic Ribosomal DNA Spacer Polymorphism Analysis

Twelve 16S-23S ribosomal DNA intergenic spacer (ITS-PCR) types were identified among 57 Staphylococcus intermedius isolates from humans and other animals. Six ITS-PCR types were host specific, and most human and canine strains belonged to the same types (A and J). Pigeon, horse, and mink strains appeared more heterogeneous