RESULTS: Unique and specific primer pairs were designed to amplify the 8 genes. The specificity of the primers was confirmed by DNA sequencing of the nanoplex PCR products and BLAST analysis. The sensitivity and specificity of V-BiA-RE nanoplex PCR assay was evaluated against the conventional culture method. The analytical sensitivity of the assay was found to be 1 ng at the DNA level while the analytical specificity was evaluated with 43 reference enterococci and non-enterococcal strains and was found to be 100%. The diagnostic accuracy was determined using 159 clinical specimens, which showed that 97% of the clinical isolates belonged to E. faecalis, of which 26% showed the HLGR genotype, but none were vancomycin resistant. The presence of an internal control in the V-BiA-RE nanoplex PCR assay helped us to rule out false negative cases.
CONCLUSION: The nanoplex PCR assay is robust and can give results within 4 hours about the 8 genes that are essential for the identification of the most common Enterococcus spp. and their antibiotic sensitivity pattern. The PCR assay developed in this study can be used as an effective surveillance tool to study the prevalence of enterococci and their antibiotic resistance pattern in hospitals and farm animals.
MATERIALS AND METHODS: This prospective study was conducted between July 2012 and December 2013. Stool samples of 59 dyspeptic patients who underwent upper endoscopy were evaluated for H. pylori stool antigen.
RESULTS: From the 59 patients who participated in this study, there were 36 (61%) males and 23 (39%) females. H. pylori was diagnosed in 24 (40.7%) gastric biopsies, 22 (91.7 %) of these being positive for the Atlas H. pylori antigen test. The sensitivity, specificity, PPV, NPV and accuracy were 91.7%, 100%, 100%, 94.6% and 96.6% respectively.
CONCLUSIONS: The Atlas H. pylori antigen test is a new non-invasive method which is simple to perform and avails reliable results in a few minutes. Thus it can be the best option for the diagnosis of H. pylori infection due to its high sensitivity and specificity.
METHODS: Recombinant PPDK (rPPDK) was expressed, purified and evaluated by Western blot. In parallel, recombinant galactose-and-N-acetyl-D-galactosamine inhibitable lectin (Gal/GalNAc lectin) was produced and tested similarly. The protein identity was confirmed by analysis using MALDI-TOF/TOF. A lateral flow dipstick (LFD) test using rPPDK was subsequently developed (rPPDK-LFD) and evaluated for serodiagnosis of ALA.
RESULTS: rPPDK was expressed as soluble protein after 4 hours of induction with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 30°C. Purification using nickel-nitrilotriacetic acid (Ni-NTA) resin yielded 1.5 mg of rPPDK from 1 L of culture with estimated molecular mass of 98 kDa on SDS-PAGE. Western blots using sera from patients with ALA, healthy individuals and other diseases probed with anti-human IgG4-HRP showed the highest sensitivity (93.3%) and specificity (100%); as compared to blots using IgG and IgG1 as secondary antibodies. Moreover, rPPDK showed better specificity when compared to rGal/GalNAc lectin. In the development of the LFD test, the optimum amount of rPPDK was 0.625 μg per dipstick and the optimum working concentration of colloidal gold conjugated anti-human IgG4 was optical density (OD) 5 (1.7 μg of anti-human IgG4). Evaluation of rPPDK-LFD using ALA patients and controls serum samples showed 87% diagnostic sensitivity and 100% specificity.
CONCLUSION: The developed rPPDK-LFD showed good potential for rapid diagnosis of ALA, and merit further multicentre validation using larger number of serum samples.
METHODS: This prospective comparative study was conducted from 2009 to 2012. Patients with ACL injuries who underwent knee arthroscopy and MRI were included in the study. Two radiologists who were blinded to the clinical history and arthroscopic findings reviewed the pre-arthroscopic MR images. The presence and type of meniscal tears on MRI and arthroscopy were recorded. Arthroscopic findings were used as the reference standard. The accuracy, sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) of MRI in the evaluation of meniscal tears were calculated.
RESULTS: A total of 65 patients (66 knees) were included. The sensitivity, specificity, accuracy, PPV, and NPV for the MRI diagnosis of lateral meniscal tears in our patients were 83, 97, 92, 96, and 90 %, respectively, whereas those for medial meniscus tears were 82, 92, 88, 82, and 88 %, respectively. There were five false-negative diagnoses of medial meniscus tears and four false-negative diagnoses of lateral meniscus tears. The majority of missed meniscus tears on MRI affected the peripheral posterior horns.
CONCLUSION: The sensitivity for diagnosing a meniscal tear was significantly higher when the tear involved more than one-third of the meniscus or the anterior horn. The sensitivity was significantly lower for tears located in the posterior horn and for vertically oriented tears. Therefore, special attention should be given to the peripheral posterior horns of the meniscus, which are common sites of injury that could be easily missed on MRI. The high NPVs obtained in this study suggest that MRI is a valuable tool prior to arthroscopy.
METHODS: Patients referred to the Endoscopic Unit for colonoscopy were recruited for the study. Stool samples were collected prior to bowel preparation, and tested for occult blood with both gFOBT and FIT. Dietary restriction was not imposed. To assess the validity of either tests or in combination to detect a neoplasm or cancer in the colon, their false positive rates, their sensitivity (true positive rate) and the specificity (true negative rate) were analyzed and compared.
RESULTS: One hundred and three patients were analysed. The sensitivity for picking up any neoplasia was 53% for FIT, 40% for gFOBT and 23.3% for the combination. The sensitivities for picking up only carcinoma were 77.8% , 66.7% and 55.5%, respectively. The specificity for excluding any neoplasia was 91.7% for FIT, 74% for gFOBT and 94.5% for a combination, whereas for excluding only carcinomas they were 84%, 73.4% and 93.6%. Of the 69 with normal colonoscopic findings, FOBT was positive in 4.3%, 23.2 %and 2.9% for FIT, gFOBT, or combination of tests respectively.
CONCLUSION: FIT is the recommended method if we are to dispense with dietary restriction in our patients because of its relatively low-false positivity and better sensitivity and specificity rates.
METHODS: Anatomical MRI and structural DTI were performed cross-sectionally on 26 normal children (newborn to 48 months old), using 1.5-T MRI. The automated processing pipeline was implemented to convert diffusion-weighted images into the NIfTI format. DTI-TK software was used to register the processed images to the ICBM DTI-81 atlas, while AFNI software was used for automated atlas-based volumes of interest (VOIs) and statistical value extraction.
RESULTS: DTI exhibited consistent grey-white matter contrast. Triphasic temporal variation of the FA and MD values was noted, with FA increasing and MD decreasing rapidly early in the first 12 months. The second phase lasted 12-24 months during which the rate of FA and MD changes was reduced. After 24 months, the FA and MD values plateaued.
CONCLUSION: DTI is a superior technique to conventional MR imaging in depicting WM maturation. The use of the automated processing pipeline provides a reliable environment for quantitative analysis of high-throughput DTI data.
KEY POINTS: Diffusion tensor imaging outperforms conventional MRI in depicting white matter maturation. • DTI will become an important clinical tool for diagnosing paediatric neurological diseases. • DTI appears especially helpful for developmental abnormalities, tumours and white matter disease. • An automated processing pipeline assists quantitative analysis of high throughput DTI data.