Detection of Helicobacter pylori cagA gene in gastric biopsies, clinical isolates and faeces.

PURPOSE: Helicobacter pylori infection is common in the developing countries. The cagA gene is a marker of pathogenicity island (PAI) in H. pylori . The aim of this study was to determine the prevalence of cagA among dyspeptic patients in Bahrain directly from gastric biopsy and stool specimen. METHODS: A total of 100 gastric biopsy samples, 16 clinical isolates and 44 faecal specimens were collected from Bahraini adult dyspeptic patients. cagA gene of H. pylori was assessed using polymerase chain reaction (PCR). RESULTS: The cagA gene was detected in 59 (59%) from biopsy specimens, 10 (62%) clinical isolates and in 10 (22.7%) faecal specimens. The detection of cagA positive H. pylori was significantly higher in patients with duodenal ulcer (80%) compared to those with other endoscopic finding (42%) (P < 0.05). CONCLUSIONS: Using PCR to detect cagA gene directly from biopsy is a rapid and reliable technique. However, using stool specimen for genotyping in our patients showed reduced sensitivity.

Antibiotic susceptibilities of Helicobacter pylori.

OBJECTIVE: The aim of this study was to evaluate the prevalence of resistance among 83 Helicobacter pylori isolates cultured from biopsies taken during routine endoscopies in 1998-1999. METHODS: Minimum Inhibitory Concentration of amoxicillin, tetracycline, clarithromycin and metronidazole were determined by Epsilometer test. RESULTS: Forty-seven strains (57%) were resistant to metronidazole, and 27 (32.5%) were resistant to clarithromycin. Twenty of the 27 strains resistant to clarithromycin were also resistant to metronidazole. None of the strains were resistant to amoxicillin or tetracycline. CONCLUSION: A high percentage of patients from Bahrain were infected with resistant strains of Helicobacter pylori. Antibiotic resistance monitoring is very important and unified national treatment policies are needed.

Rapid diagnosis of Mycobacterium tuberculosis by multiplex polymerase chain reaction from clinical specimens.

An essential element in the control of tuberculosis is the rapid, sensitive and specific identification of the causative agent. Until now, screening and diagnosis are largely based on clinical signs, radiological examination, tuberculin tests, sputum examination under the microscope, or culture for mycobacteria. Tuberculin tests lack specificity and only give an indication of previous exposure to mycobacteria. Direct microscopic examination of sputum is neither specific nor sensitive enough, and mycobacterial isolation is time-consuming. As an alternative to these classical methods, new nucleic acid-based technologies show promise as a more rapid, sensitive, and specific means of identification of mycobacteria. Two commercial standardized nucleic acid-based amplification techniques have been reported to yield reliable results within 5 to 7 hrs. Roche Amplicor MTB (Roche Diagnostic System, Somerville, N.J.) and Gen-Probe AMTB (Gen-Probe Inc., San Diego, Calif.). The amplified target is part of the 16S rRNA gene which is common to all the mycobacteria. An attempt has been made to describe the use of the target DNA, SenX3-RegX3, in a multiplex PCR to detect and differentiate M. tuberculosis from other mycobacteria directly from clinical specimens.

Chromosomal resistance to antibiotics in gonococci from Bahrain.

Ninety-one isolates of non-penicillinase-producing Neisseria gonorrhoeae from patients in Bahrain were tested for serotype, auxotype, and antibiotic susceptibility. Ten serovars and three auxotypes were found. Of the 91 isolates, 49 (54%) were serovar IB-5/7, 59 (65%) had a penicillin MIC greater than or equal to 1 mg/l, 39 (45%) had a cefuroxime MIC greater than or equal to 0.5 mg/l, and 63 (69%) had a tetracycline MIC of greater than or equal to 4 mg/l. No spectinomycin or high-level tetracycline resistance was seen. Seventy of the 91 isolates were tested against ciprofloxacin and ceftriaxone, and 40 (57%) and 26 (37%) had MICs greater than or equal to 0.03 mg/l, respectively. DNA from two penicillin-resistant isolates was capable of transforming recipient strain FA19 to donor level of penicillin and cephalosporin resistance in four steps. The first three steps were indicative of the acquisition of known resistance mutations. The existence of the fourth level transformants, with the ability of donor DNA to transform strain FA140 to higher levels of resistance, suggest the presence of another resistance mutation.

Penicillin and cephalosporin resistance in gonococci.

Non-penicillinase producing Neisseria gonorrhoeae isolated at St Mary's Hospital, London were examined for the prevalence of resistance to penicillin and for decreased susceptibility to cefuroxime. Of the 941 non-PPNG tested 100 (10.6%) were resistant to penicillin (minimum inhibitory concentration, MIC, greater than or equal to 1 mg/l) and were considered to be chromosomally-resistant N gonorrhoeae (CMRNG). Decreased susceptibility to cefuroxime (MIC, greater than or equal to 0.5 mg/l) was detected in 79% of the CMRNG. The CMRNG were also more often prototrophic and of serogroup IB than the remaining non-PPNG. The correlation coefficient for resistance to penicillin and cefuroxime was high, 0.79. Transformation experiments with both genetically-defined strains and transformants obtained using DNA from clinical isolates, showed that increased resistance to cephalosporins was acquired in three steps in close association with penicillin. We think this suggests that the loci controlling resistance to the cephalosporins are identical or closely linked to those controlling penicillin resistance.

Sampling methods for monitoring changes in gonococcal populations.

A total of 160 consecutive isolates of Neisseria gonorrhoeae was collected over a 3-month period. They were tested for their susceptibility to penicillin, erythromycin and spectinomycin and the auxotype and the serotype determined. We have evaluated two sampling methods, the collection of every fifth isolate and the first 20 isolates (10 male and 10 female) each month, to determine whether either is representative of the total population. There was no significant difference between either method of sampling and the total for detecting the predominant auxotypes and serovars or the distributions in antibiotic susceptibility. It is possible to monitor major changes in a gonococcal population, particularly susceptibility to antibiotics, using a sample of the total population.

Associations between serotype and susceptibility to antibiotics of Neisseria gonorrhoeae.

A serological classification scheme for Neisseria gonorrhoeae was used to investigate the epidemiological associations between gonococcal serotype and other bacterial and host characters. Six hundred and fifty clinical isolates of non-penicillinase producing N gonorrhoeae from the Praed Street Clinic, St Mary's Hospital, were included in this study. The strains collected represented 41 serovars, although 485 (75%) of the 650 strains belonged to five serovars. Strains of serovar IA-1/2 were commonly isolated from the cervix and tended to be sensitive to penicillin and moderately resistant to erythromycin. Strains of serovar IB-1 showed bimodal patterns of susceptibility to both penicillin and erythromycin and were obtained equally from all anatomical sites. Strains of serovar IB-2 were isolated more often from the rectum and were associated with homosexually acquired infections, whereas those of serovar IB-3 were sensitive to erythromycin and were rarely isolated from the rectum. Strains of IB-5/7 were more resistant to penicillin and erythromycin than strains of other serovars. The serological classification of N gonorrhoeae is thus a powerful tool that may be used to study biological characteristics of the gonococcus, such as susceptibility to antimicrobials and site tropism.