Rapid genotyping of genetically modified laboratory animals from whole blood samples without DNA preparation
Andrea R. HorvathPéter SánthaViktória HorváthNóra TörökI. NagyGábor JancsóCsaba VágvölgyiFerenc Somogyvári
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Duplex (building)
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Helicobacter pylori(Hp) infection can cause gastritis and peptic ulcer.The genotyping can show the essence of life.Different genotyping is correlated with different diseases and different genotyping correlated with different resistances.This article observed the Hp genotyping.There were four aspects to discuss:The correlation on different genotyping with the disease;different genotyping resistant;the different genotyping mucosal damage intensity;intervention link of traditional Chinese medicine.A brief summary of the nearly seven years of Hp genotyping research provides a new method to discover new drugs for the treatment of Hp with traditional Chinese medicine.
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Abstract Background: HIV-1 drug resistance genotyping is critical to the monitoring of antiretroviral treatment. Data on HIV-1 genotyping success rates of different laboratory specimen types from multiple sources is still scarce. Methods: In this cross-sectional study, we determined the laboratory genotyping success rates (GSR) and assessed the correlates of genotyping failure of 6837 unpaired dried blood spot (DBS) and plasma specimens from multiple studies/sources. Specimens from multiple studies in a resource-constrained setting were analysed in our laboratory between 2016 and 2019. Results: We noted an overall GSR of 65.7% and specific overall GSR for DBS and plasma of 49.8% and 85.9% respectively. The correlates of genotyping failure were viral load (VL) <10,000 copies/mL (aOR 0.3 95% CI: 0.24-0.38; p<0.0001), lack of viral load testing prior to genotyping (OR 0.85 95% CI: 0.77-0.94; p=0.002), use of DBS specimens (aOR 0.10 95% CI: 0.08-0.14; p<0.0001) and specimens from routine clinical diagnosis (aOR 1.4 95% CI: 1.10-1.75; p=0.005). Conclusions: We report rapidly decreasing HIV-1 genotyping success rates between 2016 and 2019 with increased use of DBS specimens for genotyping and note decreasing median viral loads over the years. We recommend improvement in DBS handling, viral load testing prior to genotyping and development of more sensitive assays to genotype specimens with low or undetectable viral load, especially in this era where virological suppression rates are rising due to increased antiretroviral therapy roll-out.
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Objective To establish a diagnostic method for ABO genotyping and to provide a simple and reliable technique for safe transfusion and identification of complicated blood group. Methods ABO genotyping method was developed by using multiplex-PCR-RFLP and PCR-SSP techniques. Results We developed a set of methods, which were stable and accurate for ABO genotyping. This diagnostic system were excellent for ABO genotyping after implementation. Conclusions We have established an reliable ABO genotyping diagnostic system.
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Abstract Background HIV-1 drug resistance genotyping is critical to the monitoring of antiretroviral treatment. Data on HIV-1 genotyping success rates of different laboratory specimen types from multiple sources is still scarce. Methods In this cross-sectional study, we determined the laboratory genotyping success rates (GSR) and assessed the correlates of genotyping failure of 6837 unpaired dried blood spot (DBS) and plasma specimens. Specimens from multiple studies in a resource-constrained setting were analysed in our laboratory between 2016 and 2019. Results We noted an overall GSR of 65.7% and specific overall GSR for DBS and plasma of 49.8% and 85.9% respectively. The correlates of genotyping failure were viral load (VL) < 10,000 copies/mL (aOR 0.3 95% CI: 0.24–0.38; p < 0.0001), lack of viral load testing prior to genotyping (OR 0.85 95% CI: 0.77–0.94; p = 0.002), use of DBS specimens (aOR 0.10 95% CI: 0.08–0.14; p < 0.0001) and specimens from routine clinical diagnosis (aOR 1.4 95% CI: 1.10–1.75; p = 0.005). Conclusions We report rapidly decreasing HIV-1 genotyping success rates between 2016 and 2019 with increased use of DBS specimens for genotyping and note decreasing median viral loads over the years. We recommend improvement in DBS handling, pre-genotyping viral load testing to screen samples to enhance genotyping success and the development of more sensitive assays with well-designed primers to genotype specimens with low or undetectable viral load, especially in this era where virological suppression rates are rising due to increased antiretroviral therapy roll-out.
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The use of real-time polymerase chain reaction (PCR) for detection of Strongyloides stercoralis in stool has recently been described. We compared five DNA extraction methods by using normal human stool spiked with Strongyloides ratti and tested by using a real-time PCR. The PowerSoil kit was found to be the best technique in terms of sensitivity and ease of use. The PCR detected DNA extracted from one spiked S. ratti larva diluted 10⁻². The PowerSoil kit was then used to extract DNA from 160 human survey samples. All culture positive specimens with a high and moderate larval load were identified by real-time PCR, but only 15% of specimens with low larval load were positive. Specificity was greater than 99%. The combination of the PowerSoil kit and real-time PCR reliably detected high to moderate larval numbers of S. stercoralis in stools but was less sensitive when the larval load was low.
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Abstract Background Recently, the QPLEX™ human papillomavirus (HPV) genotyping kit (QuantaMatrix, Seoul, Korea), a Microdisk™ technology-based multiplex system, was developed to detect 32 HPV genotypes. We evaluated the analytical performance of this kit by conducting a comparison study, precision evaluation and interference testing. Methods A total of 1594 cervical swab specimens were used to compare the QPLEX™ HPV genotyping kit with other commercially available kits (GeneFinder HPV Liquid Bead MicroArray Genotype polymerase chain reaction [PCR] kit, Infopia, Seoul, Korea; PANArray™ HPV Genotyping Chip, PANAGENE, Daejeon, Korea). For the determination of precision, we evaluated four types of precision profiles: repeatability, lot-to-lot variability, operator-to-operator variability and site-to-site variability. In addition, interference tests were performed with various interferents. Results The results of the QPLEX™ HPV genotyping kit showed almost perfect agreement with the other commercially available HPV genotyping assays. The combined precision was acceptable. In addition, there was no tested interferent that affected the results of the QPLEX™ HPV genotyping kit. Conclusions The QPLEX™ HPV genotyping kit showed acceptable analytical performance in our study. This assay could be a suitable option for HPV genotyping in routine and follow-up tests.
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