Next generation sequencing to determine the cystic fibrosis mutation spectrum in Palestinian population.

An extensive molecular analysis of the CF transmembrane regulator (CFTR) gene was performed to establish the CFTR mutation spectrum and frequencies in the Palestinian population, which can be considered as an understudied population. We used a targeted Next Generation Sequencing approach to sequence the entire coding region and the adjacent sequences of the CFTR gene combined with MLPA analysis of 60 unrelated CF patients. Eighteen different CF-causing mutations, including one previously undescribed mutation p.(Gly1265Arg), were identified. The overall detection rate is up to 67%, and when we consider only CF patients with sweat chloride concentrations >70 mEq/L, we even have a pickup rate of 92%. Whereas p.(Phe508del) is the most frequent allele (35% of the positive cases), 3 other mutations c.2988+1Kbdel8.6Kb, c.1393-1G>A, and p.(Gly85Glu) showed frequencies higher than 5% and a total of 9 mutations account for 84% of the mutations. This limited spectrum of CF mutations is in agreement with the homozygous ethnic origin of the Palestinian population. The relative large portion of patients without a mutation is most likely due to clinical misdiagnosis. Our results will be important in the development of an adequate molecular diagnostic test for CF in Palestine.

Isolation and characterisation of Shiga toxigenic Escherichia coli strains from northern Palestine.

Shiga toxigenic Escherichia coli (STEC) isolates from symptomatic and asymptomatic patients in northern Palestine in 1999 were screened for serotype O157 and characterised for virulence genes by multiplex PCR assay. Of the 176 STEC isolates, 124 (70.5%) were of serotype O157. All these isolates carried the gene for Shiga toxin type 1 (stx,) and 112 (90.3%) carried stx2. The intimin encoding gene locus eae was detected in 16 isolates (12.9%) and the enterohaemolysin encoding gene, hlyA, in 18 (14.5%). Statistical analysis showed a significant association between the presence of eaeA and hlyA, either alone or combined with stx1 and stx2 genes in O157 isolates from symptomatic infection. ERIC-PCR analysis of DNA from 80 serotype O157 isolates revealed three major clonal populations.

Screening of some Palestinian medicinal plants for antibacterial activity.

Antibacterial activity of organic and aqueous extracts of 15 Palestinian medicinal plants were carried against eight different species of bacteria: Bacillus subtilis, two Escherichia coli species, Staphylococcus aureus (methicillin resistant), two S. aureus (methicillin sensitive) species, Pseudomonas aeruginosa, and Enterococcus fecalis. Of the 15 plants tested, eight showed antibacterial activity. Each plant species has unique against different bacteria. The most active antibacterial plants against both gram-positive and gram-negative bacteria were Thymus vulgaris and Thymus origanium. The organic and aqueous extract from the same plants showed different activities; the organic extract showed the same or greater activity than the aqueous extract. Finally, the hole-plate diffusion method showed larger activity than the disc diffusion method.

Molecular, antibiogram and serological typing of Staphylococcus aureus isolates recovered from Al-Makased Hospital in East Jerusalem.

Staphylococcus aureus is a major cause of nosocomial infections and a risk in patients who have either undergone surgery or are on haemodialysis. The S. aureus infections in patients admitted to the clinical departments of Al-Makased Charitable Hospital in Jerusalem during a period of one year were investigated. Isolates included were from blood, surgical wounds, or other nonsuperficial sites. Of 63 isolates available for analysis, 46 (73.0%) expressed type 8 capsular polysaccharide; 13 (20.7%), type 5 capsular polysaccharide; only 4 isolates (6.3%) did not express type 5 or type 8 antibodies. The strains fitted in 7 different antibiogram types, with the type showing resistance only to penicillin and ampicillin prevalent in 34 out of 63 isolates (54.0%). Of the 12 methicillin-resistant S. aureus (MRSA) isolates (19.1%), 8(66.7%) possessed the type 8 capsule and 4(33.7%) the type 5 capsule. Pulsed-field gel electrophoresis of all isolates with the restriction-endonuclease enzymes Sma I revealed 34 patterns demonstrating that no single methicillin-sensitive S. aureus strain was endemic in the hospital. However, all MRSA isolates with a type 8 capsule showed identical PFGE patterns using the 2 restriction-endonuclease enzymes Sma I and SST II. Moreover, type 5 isolates showed identical patterns (one isolate differed from the rest with one band only). These data suggest and confirm the clonality of type 5 and type 8 MRSA isolates. Analysing the results of the capsular and antibiogram typing schemes in conjunction proved useful and suggested that such an analysis can be employed as a helpful epidemiological tool in hospitals with limited resources.