Malaria, caused by the parasite Plasmodium and transmitted by mosquitoes, is an epidemic that mainly occurs in tropical and subtropical regions. As treatments differ across species of malarial parasites, there is a need to develop rapid diagnostic methods to differentiate malarial species. Herein, we developed a multiplex malaria Pan/Pf/Pv/actin beta loop-mediated isothermal amplification (LAMP) to diagnose Plasmodium spp., P. falciparum, and P. vivax, as well as the internal control (IC), within 40 min. The detection limits of the multiplex malaria Pan/Pf/Pv/IC LAMP were 1 × 102, 1 × 102, 1 × 102, and 1 × 103 copies/µL for four vectors, including the 18S rRNA gene (Plasmodium spp.), lactate dehydrogenase gene (P. falciparum), 16S rRNA gene (P. vivax), and human actin beta gene (IC), respectively. The performance of the LAMP assay was compared and evaluated by evaluating 208 clinical samples (118 positive and 90 negative samples) with the commercial RealStar® Malaria S&T PCR Kit 1.0. The developed multiplex malaria Pan/Pf/Pv/IC LAMP assay showed comparable sensitivity (100%) and specificity (100%) with the commercial RealStar® Malaria S&T PCR Kit 1.0 (100%). These results suggest that the multiplex malaria Pan/Pf/Pv/IC LAMP could be used as a point-of-care molecular diagnostic test for malaria.
Background: Cerebrospinal fluid (CSF) analysis is essential for diagnosing various disorders affecting the central nervous system (CNS). Traditionally, CSF cell count analysis is performed manually using a Neubauer chamber hemocytometer, which is labor-intensive and prone to subjective interpretation. Methods: In this study, we evaluated the analytical and clinical performance of the Microscanner C3, an automated cell counting system, for CSF analysis using artificially prepared samples and 150 clinical CSF samples. Results: The lowest detectable white blood cell (WBC) count was 3.33 cells/µL, and the lowest detectable red blood cell (RBC) count was 3.67 cells/µL. The coefficients of variation (CV%) for the Microscanner C3 were lower than those for the Neubauer chamber at all cell concentrations. The correlation coefficients (R) between the Microscanner C3 and conventional methods were high: 0.9377 for WBCs and 0.9952 for RBCs when compared with the Neubauer chamber, and 0.8782 for WBCs and 0.9759 for RBCs when compared with the flow cytometer. Additionally, the Microscanner C3 showed good agreement with both the Neubauer chamber and flow cytometer in the Passing–Bablok regression analysis and Bland–Altman analysis for WBC count at all concentrations and RBC count at concentrations of 0–1000 cells/µL. Conclusions: The Microscanner C3 proved to be more sensitive, precise, and consistent compared to the conventional hemocytometer. The new system is also compact, convenient, and cost-effective, making it a valuable option for clinical laboratories.
A pre-transfusion crossmatch test is crucial for ensuring safe blood transfusions by identifying the compatibility between donor and recipient blood samples. Conventional tube methods for crossmatching have limitations, including subjectivity in result interpretation and the potential for human error. In this study, we evaluated the diagnostic performance of a new crossmatch test using Microscanner C3, which can overcome these shortcomings. The crossmatch test results using the method were obtained in 323 clinical samples. The sensitivity, specificity, positive predictive value, negative predictive value, and concordance rate of the crossmatch test using Microscanner C3 were 98.20%, 100.00%, 100.00%, 98.11%, and 99.07%, respectively. The diagnostic performance of the new system offers a promising alternative to conventional tube methods for pre-transfusion crossmatch testing. Microscanner C3 could also increase the automation, standardization, and accuracy of crossmatch tests. The crossmatch test using Microscanner C3 is thought to increase the efficiency and reliability in identifying blood samples suitable for transfusion, thereby improving patient safety and optimizing the use of blood products in clinical settings.