Comparison of multiplex reverse transcription-PCR-enzyme hybridization assay with immunofluorescence techniques for the detection of four viral respiratory pathogens in pediatric community acquired pneumonia

The burden of illness due to viral respiratory pathogens in the pediatric population is increasingly being recognized. Children are considered among the groups at highest risk for viral pneumonia-associated morbidity and mortality. Clinical discrimination between different causative agents is extremely difficult. The main problems have been the lack of ‘gold standard’ method for obtaining viral etiology (Liolios et al., 2001). We believe that the identification of these viruses as causes of respiratory disease in these patients is the first step in determining how frequently they may cause serious problems and, hence, how hard we should push with accepted treatments. In this study, our aim was to compare between two modalities for diagnosis of viral illness among children with community-acquired pneumonia (CAP). Multiplex reverse transcription-PCR-enzyme hybridization assay and immunofluorescence antigen detection techniques for the detection of four viral respiratory pathogens (Influenza viruses A & B and Respiratory Syncitial Viruses A & B) were targeted to evaluate their diagnostic yield for these patients in our study. Among 56 respiratory samples were evaluated from children with clinical and radiological criteria of CAP; twenty-one patients had viral pneumonia proved by multiplex RT-PCR and/or IF technique with disease prevalence 35% (95% CI: 23:49). All 21 specimens were positive by multiplex RT-PCR, while 20 out of them were positive by IF. All results showed no discordance of detected viral pathogen. Initial comparison of IF results to those of RT-PCR generated a sensitivity 100% (95% CI: 83:100), a specificity 97.2% (95% CI: 85:99.9), a positive predictive value 95% (95% CI: 23:49.6), and a negative predictive value of 100% (95% CI: 74:99).Conclusion: Multiplex reverse transcription PCR has an excellent potentials for diagnosis of viral pneumonia with a cost effective advantage in assessing simultaneously multiple clinically significant viruses. Rapid antigen tests for diagnosis of variable respiratory viruses, can be useful in etiological diagnosis of community acquired lower respiratory tract infection as well specially with the proved high sensitivity and predectivity in our study

Serum YKL-40 and assessment of severity of bronchial asthma in Egyptian children

Background: Serum and lung tissue levels of a chitinase-like protein YKL-40 have recently been found to be increased in patients with bronchial asthma. Furthermore, serum YKL-40 levels correlated positively with thickening of the lung sub-epithelial basement membrane, frequency of rescue inhaler use, and deterioration in pulmonary function in European asthmatic subjects. Objectives: to assess the role of YKL-40 measurement in evaluating asthma severity, compared to clinical assessment and the related pulmonary function tests. Methods: We quantified serum YKL-40 levels in two groups of Egyptian asthmatics: One group with mild to moderate asthma, and one with severe asthma. Serum YKL-40 was measured by enzyme-linked immunosorbent assay (ELISA) kits (Quidel). Clinical scoring of asthma severity by Pediatric Asthma Score (PAS) and pulmonary functions were performed. Results: The serum levels of YKL-40 were significantly elevated in severely asthmatic Egyptian children compared with the other group (151ng/ml- 72ng/ml; p < 05). YKL-40 levels were correlated positively to PAS (r=0.34, p < 0.05), and inversely to FEV1 (r= -0.32, p < 0.5). Best cut off value of YKL-40 for asthma prognosis was 90 ng/ml, sensitivity 86.5%, specificity 81%, and diagnostic accuracy of 85%. Conclusions: YKL-40 is found in increased quantities in the sera of severe asthmatics, and correlated significantly to PAS and pulmonary function deterioration. YKL-40 is considered a promising biomarker for asthma severity and pulmonary remodeling warranting further study as a potential novel pathway to disease management. Keywords: YKL-40, Asthma, severity, Egyptian, Children, biomarkerEgypt J Pediatr Allergy Immunol 2011;9(2):93-9

Multiplex polymerase chain reaction: Could change diagnosis of Ventilator-associated pneumonia in pediatric critical care units to the fast track?

Background: Ventilator-associated pneumonia (VAP) is a frequent hospital-acquired infection in critically ill children. The increasing incidence of infections by antibiotic-resistant pathogens adds significantly to the cost of hospital care and to the length of hospital stays. Besides clinical prerequisites for presumptive diagnosis of VAP, rapid identification of the causative pathogen is essential for appropriate treatment. Aim of study: To identify the causative bacterial pathogens of VAP by both conventional microbiological cultures and multiplex reverse transcriptase reaction (m-PCR) methods with assessment of turnaround time for both diagnostic modalities together with their diagnostic accuracy. Methods: Patients were diagnosed to have VAP when their Clinical Pulmonary Infection Score (CPIS) index was more than 6. Endotracheal aspirate was subjected to both microbiological cultures and multiplex PCR for bacterial pathogens. Results: Multiplex-PCR showed better sensitivity and positive predictive value than bacterial culture for etiological diagnosis of VAP. Acinetobacter and Klebsiella pneumoniae were the most common identified pathogens. Mean turnaround times were 6 h for multiplex PCR and 72 h for conventional microbiology. Significant shorter turnaround time was recorded with m PCR compared to microbiological culture. Conclusion: Multiplex-PCR permits simultaneous detection of several bacterial pathogens in a single reaction with best turnaround time that permit optimization of emergency diagnosis of VAP and subsequently improve early management of selective bacterial pathogens. Keywords: m-PCR, Ventilator-associated pneumonia, Bacterial diagnosis, Turnaround tim

Egyptian consensus on treat-to-target approach for osteoporosis: a clinical practice guideline from the Egyptian Academy of bone health and metabolic bone diseases

Abstract Background This study was carried out to achieve an Egyptian expert consensus on a treat-to-target management strategy for osteoporosis using Delphi technique. A scientific committee identified researchers and clinicians with expertise in osteoporosis in Egypt. Delphi process was implemented (2 rounds) to establish a consensus on 15 clinical standards: (1) concept, (2) diagnosis, (3) case identification, (4) whom to treat, (5) who should treat?, (6) case stratification and intervention thresholds, (7) falls risk, (8) investigations, (9) treatment target, (10) management, (11) optimum treatment duration, (12) monitoring, (13) drug holiday, (14) osteoporosis in men, and (15) post-fracture care and fracture liaison service. Results The surveys were sent to an expert panel (n = 25), of whom 24 participated in the two rounds. Respondents were drawn from different governorates and health centres across Egypt including the Ministry of Health. Most of the participants were rheumatologists (76%), followed by internists (8%), orthopaedic doctors (4%), rehabilitation doctors (4%), primary care (4%), and ortho-geriatrics (4%) physicians. Seventy-two recommendations, categorised into 15 sections, were obtained. Agreement with the recommendations (rank 7–9) ranged from 83.4 to 100%. Consensus was reached (i.e. ≥ 75% of respondents strongly agreed or agreed) on the wording of all 15 clinical standards identified by the scientific committee. An algorithm for the management of postmenopausal osteoporosis has been suggested. Conclusion A wide and representative panel of experts established a consensus regarding the management of osteoporosis in Egypt. The developed guidelines provide a comprehensive approach to the assessment and management of osteoporosis for all Egyptian healthcare professionals who are involved in its management