Quality assurance program for molecular medicine laboratories

Molecular diagnostic methods have played and continuing to have a critical role in clinical laboratories in recent years. Therefore, standardization is an evolutionary process that needs to be upgrade with increasing scientific knowledge, improvement of the instruments and techniques. The aim of this study was to design a quality assurance program in order to have similar conditions for all medical laboratories engaging with molecular tests. We had to design a plan for all four elements; required space conditions, equipments, training, and basic guidelines. Necessary guidelines was prepared and confirmed by the launched specific committee at the Health Reference Laboratory. Several workshops were also held for medical laboratories directors and staffs, quality control manager of molecular companies, directors and nominees from universities. Accreditation of equipments and molecular material was followed parallel with rest of program. Now we are going to accredit medical laboratories and to evaluate the success of the program. Accreditation of medical laboratory will be succeeding if its basic elements are provided in advance. Professional practice guidelines, holding training and performing accreditation the molecular materials and equipments ensured us that laboratories are aware of best practices, proper interpretation, limitations of techniques, and technical issues. Now, active external auditing can improve the applied laboratory conditions toward the defined standard level

Knowledge, attitude and practice of nurses toward peak expiratory flow meter in primary health care centers in Kuwait

Peak expiratory flow meter [PEFM] is an easy to use, relatively cheap device that can be used for guiding management of bronchial asthma by the patients at home according to a preset plan by health care workers. The aim of the study is to reveal the extent of knowledge and perception of nurses about PEFM and factors affecting their knowledge. Out of the total primary health care centers in Kuwait, 50% were randomly selected. Out of 699 nurses currently working in the selected centers, 516 nurses were interviewed for this study with an overall response rate of 73.8%. The results of this study showed that nurses had a relatively low total knowledge score percent of 64.7 +/- 7.3%. The lowest individual mean percent score was that of procedures and steps of measuring peak expiratory flow rate [39.0 +/- 24.1%]. The highest percent knowledge score was that of benefits of use and content instructions for teaching patients [78.3 +/- 19.5% and 78.1 +/- 12.0%, respectively]. Sociodemographic factors did not affect the total knowledge score. Receiving training, availability of PRFM in the health center and being responsible about taking the measurements for patients proved to significantly affect the level of knowledge of nurses. Training nurses about use of PEFM and providing primary health care centers with the devices would plan an important role to improve knowledge of nurses and hence improve domestic health care of patients with obstructive lung diseases

Genotyping pattern among Iranian HCV positive patients

Successful treatment to eliminate HCV RNA depends on the identified genotype. In the present study, we compared the frequency of different HCV genotypes, during four years study [2004 till 2008]. Sera specimens were received from 16 provinces of Iran. We used High Pure Viral Nucleic Acid Purification kit for extraction and samples were tested with improved form of RT-PCR technique. HCV genotypes were determined using Amplisense PCR kit and Amplicor HCV Monitoring Version 2 test utilized a reverse transcription [RT]-PCR approach to quantitative HCV RNA. Two hundreds six HCV positive specimens were entered to the study out of 389 tested samples. Type 3a was the most frequent type [46.6%], followed by type 1 [including 1a and 1b with 25.73% and 17.47% for each respectively] with 43.2%. Looking through collected results of the four years study confirmed the rate of HCV infection in those single genotypes 1b, 3a were slightly increased from 12.22% and 38.88% in the first year to 18.66 and 46.51% in the fourth year of the study period. The analyzed data proved that some patients were infected with two different types. High viral load was also more correlated to genotype 1 than other types

Specific PCR assay for rapid and direct detection of Neisseria meningitidis in cerebrospinal fluid specimens

Neisseria meninigitidis is one of the most frequently encountered microorganisms associated with central nervous system infections. The aim of this study was to evaluate a PCR-based assay for specific and rapid detection of N. meninigitidis in CSF specimens. Since April 2002 to July 2006, 130 CSF specimens were collected from patients suspected of having baterial meningitis. Bacterial isolation and identification was carried out according to the standard bacteriological methods. The PCR was used to amplify a 101bp fragment of capsular transport gene A [ctr A] of N. meningitidis. PCR yielded an amplified product with the expected size of 101 base pair fragment. Sensitivity test proved 500 mg of N. meningitidis DNA as the final detection limit and specificity test revealed no cross-reaction for a wide range of respiratory pathogenic organisms. The PCR assay was more sensitive than the bacterial culturing. It might be possible to apply this procedure for rapid diagnosis of meningococci in clinical samples

Incidence and potential risk factors of low birth weight among full term deliveries

Low birth weight [less than 2500 grams] [LBW] is a reliable indicator in monitoring and evaluating the success of maternal and child health programs. Giving birth to a LBW infant is influenced by several factors. The aim of the study was to determine the incidence of LBW among live born full term deliveries in Al-Adan governmental hospital and to study the possible determinants of its occurrence. The first phase of the study was a descriptive one including all live born full term deliveries of women attended the hospital within the first 2 months of pregnancy and followed up till delivery. In the second phase LBW women [cases] were compared with a double number of women who gave birth to normal birth weight infants. Among 939 women eligible for the study, the incidence of LBW was 7.8%. Reproductive age lower than 25 years, gestational age lower than 40 weeks, history of previous abortion, primiparity, maternal underweight and anemia were detected as independent risk factors for LBW. On the other hand, obesity was a protective factor. Low birth weight occurs frequently in Kuwait, although its incidence is much lower than in many countries in the region. Prenatal management of modifiable factors and adequate antenatal care and screening for susceptible women should be a target for all obstetricians for reducing the incidence of LBW

Prevalence of hepatitis C virus among out patients of a private laboratory in Tehran

With an estimated 170 million infected individuals, hepatitis C virus [HCV] has a major impact on public health. Frequency of HCV infection was studied in 785 referred patients to a private Laboratory, Tehran, Iran from April 2003 until March 2006 by PCR method. Analyzed results revealed infection rate was 40.27%, 35.51% and 33.09% in the first, second and third year, respectively. Frequency rate of the infection shows a fluctuated shape during months of three years. Maximum rate of the HCV infection was in May of the first and in June of the second and third year, respectively. It shows decrease and increase within rest months in each year. HCV infection rate was higher in 31 to 40 years old group of patients. Analysis of the data revealed higher HCV infection in men than women had. It was concluded that HCV frequency pattern underlines specific attention to suspected patients during high rate time

[Serotyping of Neisseria meningitides in conscripts with meningitis admitted to five military hospital in Tehran between September 2004 and September 2006]

Background: Meningococcal meningitis is one of the most feared disorders with high rate morbidity and mortality. There is no exact information of meningococcal meningitis incidence in military personnel. Obligate vaccination of the military with meningococcal vaccine is a strategy with the potency to provide active protection of the conscripts before being dispatched to training centers. Although, sporadic cases of meningitis in conscripts occurred. The aim of this study was serotyping of the bacterium; Neisseria meningitides isolated from patients with meningitis which admitted to five military Hospitals

Materials and methods: In this cross- sectional study for determination of meningococcal meningitis serotypes in conscripts submit to five military hospitals the investigation was designed and carried out from September 2003 to September 2006. In this period, 12 cerebrospinal fluids of conscripts with clinical signs and symptoms of meningitis were collected, and the laboratory tests were done. In this study enriched Thayer Martin medium and standard bacteriological methods were used. The sample without bacterial cells or PMNs were centrifuged [10000 xg for 5 min]. The precipitate was cultured and direct smear was performed. Isolated bacterial strains was recognized by biochemical tests, and Neisseria meningitides strains were serotyped by specific antiserum

Results: During the study, 12 cases of meningitis in conscripts were seen. Neisseria meningitides was isolated only from 6 patients. Serotyping analysis revealed that 5 strains belong to Serotype C and 1 strain was to serotype B. Neisseria sica was isolated from one patient. In three patients there was no bacteriological evidence. In two patients, consumption of antibiotic before lumbar puncture results to no bacterial growth, but direct smear showed gram negative diplococci. In one patient recurrent meningococcal infection was occurred. Complement components analysis revealed deficiency in C3, C4 and CH50 [the rate was below 80 mg per deciliter]

Conclusions: the results of this study showed that, Neisseria meningitides was isolated from six patients which are all conscripts. It is necessary to find the reasons that why vaccinated conscripts infected by Neisseria meningitides serotype C. Further analysis showed that, five patients had complement deficiency and further research is necessary. However, 6 Neisseria meningitides strains were isolated from meningococcal vaccinated conscripts around the country [Tehran, Shiraz, Yazd and Oslavieh] in the period of study. Based on the findings of this study, effectiveness of the vaccine was desirable and the disease had been controlled in this population group. Because of complements deficiency in patients with meningococcal meningitis, other preventing methods for eradication of disease are necessary

Serotyping of Neisseria meningitides in conscripts with meningitis admitted to five military hospital in Tehran between September 2004 and September 2006

Meningococcal meningitis is one of the most feared disorders with high rate morbidity and mortality. There is no exact information of meningococcal meningitis incidence in military personnel. Obligate vaccination of the military with meningococcal vaccine is a strategy with the potency to provide active protection of the conscripts before being dispatched to training centers. Although sporadic cases of meningitis in conscripts occurred The aim of this study was serotyping of the bacterium; Neisseria meningitides isolated from patients with meningitis which admited to five military Hospitals. In this cross- sectional study for determination of meningococcal meningitis serotypes in conscripts submit to five military hospitals the investigation was designed and carried out from September 2003 to September 2006. In this period, 12 cerebrospinal fluids of conscripts with clinical signs and symptoms of meningitis were collected, and the laboratory tests were done. In this study enriched Thayer Martin medium and standard bacteriological methods were used. The sample without bacterial cells or PMNs were centrifuged [10000 xg for 5 min], The precipitate was cultured and direct smear was performed.Isolated bacterial strains was recognized by biochemical tests, and Neisseria meningitides strains were serotyped by specific antiserum. During the study, 12 cases of meningitis in conscripts were seen. Neisseria meningitides was isolated only from 6 patients. Serotyping analysis revealed that 5 strains belong to Serotype C and 1 strain was to serotype B. Neisseria sica was isolated from one patient. In three patients there was no bacteriological evidence. In two patients, consumption of antibiotic before lumbar puncture results to no bacterial growth, but direct smear showed gram negative diplococci. In one patient recurrent meningococcal infection has occurred. Complement components analysis revealed deficiency in C3, C4 and CH50 [the rate was below 80 mg per deciliter]. The results of this study showed that, Neisseria meningitides was isolated from six patients which are all conscripts. It is necessary to find the reasons that why vaccinated conscripts infected by Neisseria meningitides serotype C. Further analysis showed that, five patients had complement deficiency and further research is necessary. However, 6 Neisseria meningitides strains were isolated from meningococcal vaccinated conscripts around the country [Tehran, Shiraz, Yazd and Oslavieh] in the period of study. Based on the findings of this study, effectiveness of the vaccine was desirable and the disease had been controlled in this population group. Because of complements deficiency in patients with meningococcal meningitis, other preventing methods for eradication of disease are necessary

Efficiency of PCR method for screening pulmonary tuberculosis patients under DOTs protocol therapy

Tuberculose has increased in the recent years. Use of a sensitive screening method, can be one of the influential parameters in reduction of pulmonary tuberculosis. Current screening method in the laboratories is based on observation of bacilli in stained smear by Ziel-Neelsen procedure. The aim of this research was to study the PCR efficiency in all confirmed patients of Hamadan province. Twenty eight patients were registered as having pulmonary tuberculosis by the tuberculosis comity of the Hamadan province. All these patients had positive results for staining and culture. Amonst these registered patients, we could access only to the 12 patients during the study. Sputums were collected from early stage of diagnosis and continued during treatment at the end of each month. All these patients were under the treatment by the DOTs protocols. All samples were tested by PCR and the results were compared with Ziehl-Neelsen staining method that is recommended procedure by WHO. Compared results showed none of the samples were positive by staining method at the end of second month up to the last month of treatment, while PCR changed to negative gradually. It was negative in 5 and 4 patients at the end of the third and forth month, respectively. Specimens of the three remanied patients were continuously positive up to the end of treatment period. Gradually changing PCR results to negative in three forth of studied patients means it can be applicable screening tools, but one forth remained positive cases needs more study for the evaluation of target gene role and efficiency of the test

Comparison of four PCR tests for the detection of mycoplasma pneumoniae

Recently, PCR is being used more frequently as a diagnostic method to detect M. pneumoniae. We used primer pairs reported by Van Kuppeveld, Leng, Lunerberg, and Bernet targeting 16s rRNA, P1 protein, tuf genes, and a short DNA sequence [MP5] to evaluate the sensitivity among different PCRs. Reoptimization experiments showed that tuf PCR had the highest sensitivity amongst these four PCRs, detecting 10 organisms. Detection limit for the rest of the PCRs was 100 copies of DNA. This study confirmed that 92°C would be the best dissociation temperature rather than higher temperatures that are still being used frequently in other studies. Besides, accurate optimizing of the annealing temperature and extension time had important roles on the sensitivity as well as using milli-Q distilled water rather than double distilled water. Experiments done on MP5 PCR proved that the non-specific products mentioned in previous studies were not eliminated by increasing the annealing temperature, although they disappeared on gel electrophoresis after careful optimization of extension time