Various molecular methods have been used for the rapid identification of mycobacterial species. In this survey, evaluation of antibiotic resistance and PCR-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene was carried out for identification of non-tuberculosis mycobacteria (NTM) isolates from different clinical specimens. Forty-eight different mycobacterial isolates were selected and followed by the conventional and PRA of hsp65 for species identification. The antibiotic susceptibility test was carried out according to standard methods. A 439 bp PCR product of hsp65 in all selected isolates was amplified and digested with the BstEII and HaeIII restriction enzymes. The restriction fragment length polymorphism (RFLP) patterns were analyzed for species identification. Using PRA for 48 mycobacterial selected isolates, including 15 M. tuberculosis, one M. bovis and all 32 isolates of NTM, revealed 11 different species among the NTM isolates. The most frequent NTM isolates were M. kansasii, M. gordonae III, M. marinum, M. chelonae, M. scrofluaceum and M. gastri. In most cases, the PRA results were perfectly in accordance with the classical biochemical method. Combination of resistance to rifampin and isoniazid was present among M. kansasi, M. gordoniae III, M. scrofluaceum, M. chelonae, M. marinum, M. gastri, M. gordoniae II and M. trivale isolates. A high incidence of co-resistance to six, five, four and three anti-TB drugs was observed in 18.5%, 9.1%, 6.6% and 11.7% of all NTM isolates, respectively. Our results showed that PRA, in comparison with classical methods, is rapid and accurate enough for the identification of mycobacterial species from LJ medium. Additionally, we found that in Iran we have a highly diverse population of NTM isolates among patients suspected of having TB.

ÖZETİran sınır bölgesindeki pulmoner tüberküloz izolatlarında yüksek düzeyde izoniazid direnci katalaz-peroksidazı kodlayan katG'deki multipl mutasyonla koreledir Bu çalışmanın amacı, katG

medical or immunocompromising conditions. Clinical and epidemiological data were recorded and respiratory samples including nasopharyngeal aspirate or nasopharyngeal swabs were obtained from all children less than 14 years old with acute respiratory tract infections. HBoV was screened in all respiratory samples by real time PCR, in addition to 13 common respiratory viruses. During the study, HBoV was detected in respiratory samples from 25 (2%) of 1016 symptomatic patient. HBoV coexistence with other respiratory pathogens occurred in 72% (18/25) of respiratory samples from symptomatic patients. HBoV infections were detected in every month except June and July with peaks in the month of September, October, November, and December. The main diagnosis in 13 patients (52%) with HBoV was radiologically confirmed pneumonia. For the other 12 patients with HBoV infections the main diagnosis were gastroenteritis(4 cases), chest exacerbation (3 cases), upper respiratory tract infections (2 cases), persistent fever (1 case), seizure (1 case), otitis media (1 case). The main clinical signs and symptoms of HBoV positive patients included fever, cough tachypnea, dyspnea, crackles, wheezing, abdominal pain, vomiting and diarrhea. The present study suggest that HBoV may be a fairly common cause of pneumonia in high-risk children hospitalized with acute respiratory infections and associated with morbidity. However, further study is needed to clarify if HBoV plays a pathogenic role in community acquired pneumonia in high-risk children.

Background and Aims Non-structural protein 4 (NSP4), encoded by group A rotavirus (RVA) genome segment 10, is the first recognized virus-encoded enterotoxin. Recently, a new classification system for RVAs was proposed and a total of 14 NSP4 genotypes (E1 to E14) are currently described. Methods A total of 1391 faecal specimens collected from children under 5 years old were screened by ELISA for the presence of RVA antigen. NSP4-encoding genes of RVA positive strains were analyzed using a semi-nested RT-PCR. Results Genotypes E1 and E2 were identified in 183 (70.1%) and 78 (29.9%) samples, respectively. This report represents the first investigation on the genetic diversity of RVA NSP4 genes in Tunisia. Tunisian RVA strains analysed in the present study belonged to 2 different genotypes: E1 and E2. Such a result is concordant with literature data: indeed, although 14 RV NSP4 genotypes have been identified to date, previous molecular characterization has shown that most of the diversity in the NSP4-encoding gene lies in genotypes E1 and E2. Other studies, however, have detected unusual strains carrying genotypes E3 and E13. Moreover, a predominance of NSP4 genotype E1 was observed over the entire period of study, from 2006 to 2008. Such a result was also quite expected as previous investigations have also shown that NSP4 genotype E1 was largely predominant among children worldwide. Conclusions These results underline the need for further investigations to assess the validity of NSP4 as a suitable target for epidemiologic surveillance of rotavirus infections and vaccine development.