Purpose: Describing the utility of Polymerase Chain Reaction for Cytomegalovirus (CMV-PCR) in the diagnosis of suspected viral anterior uveitis (AU). Patients and methods: We analyzed aqueous humor (AH) CMV-PCR positivity and treatment modifications in 47 eyes of 46 patients with viral uveitis and its correlation with high intraocular pressure (HIOP), uveitis clinical characteristics and time intervals from samples to uveitis diagnosis and to relapse. Results: CMV-PCR positive results occurred in 13 eyes (27.7%) of 12 patients. They were more frequent in HIOP eyes (34.2%, p = 0.047) and with Posner–Schlossman Syndrome (56.2%, p = 0.002). CMV-PCR positivity (p = 0.001) and HIOP (p = 0.038) increased the probability of treatment change. Although CMV-PCR positive results decreased over time (p = 0.002), they were not related to activity or proximity to inflammatory uveitis episode. Conclusion: HIOP AU eyes should be considered for CMV-PCR AH analysis due to possible treatment modifications.

Background: Nigeria accounts for the greatest burden of malaria disease globally. Malaria control requires an effective treatment after diagnosis. The efficacy of antimalarial drugs can be assessed through the analysis of genetic changes associated with reduced drug sensitivity. Methods: This study includes the analysis of the markers associated with artemisinin (pfk13), sulfadoxine–pyrimethamine (pfdhfr and pfdhps), and chloroquine and its derivatives (pfmdr1 and pfcrt) resistances, in blood samples collected from asymptomatic children in south-western Nigeria. Results: The 25.95% of samples showed a number of mutations in pfk13 gene. Among those, the validated, C580Y, and the candidate, R515K, mutations by WHO were detected. Twenty-seven pfdhps different haplotypes were observed, with the haplotype ISGKAA as the most prevalent (18.80%), followed by IFGKAA (12.78%) and IAGKAA (11.28%). The VAGKGS was the most common haplotype carrying the I431V mutation (10.53%). Combinations of alleles in pfdhfr and pfdhps genes provided a 40.98% of samples with the partially resistant haplotype (IRNG). No samples exhibited the ‘fully resistant’ or ‘super resistant’ pfdhprf–pfdhps combinations, but one sample contained mutations at pfdhfr 51I, 59R, and 108N with pfdhps 431V, 436A, A437G and 540E. The analysis of pfcrt 72–76 variants disclosed a 12.12% of samples with the mutant-type (CVIET). No double mutant pfmdr1 haplotypes 86Y/1246Y (YY) were detected, nor was the haplotype formed by the alleles 86Y pfmdr1 + pfcrt 76 T (YT). Conclusions: There was no evidence of parasite genomes harbouring multilocus mutations conferring multidrug resistance, although evidence of a validated (C580Y) and a candidate (R515K) mutation in pfk13 gene, high frequency pfdhfr mutant alleles and high variability of pfdhps haplotypes were found in this study, which provides a baseline information essential in monitoring P. falciparum resistances.

: Malaria control and elimination require prompt diagnosis and accurate treatment. Conventional methods such as rapid diagnostic tests (RDTs) and microscopy lack the characteristics to detect low parasitemias, commonly found in asymptomatic parasitemias and/or submicroscopic malaria carriers. On the contrary, molecular methods have higher sensitivity and specificity. This study evaluated the performance of two commercial real-time polymerase chain reaction (PCR) assays, RealStar^® Malaria PCR (RealStar-genus) and RealStar Malaria Screen&Type PCR (RealStar-species), compared with the reference Nested Multiplex Malaria PCR, for the detection of the main five Plasmodium species affecting humans. A total of 121 samples were evaluated. Values of sensitivity (98.9% and 97.8%) and specificity (100% and 96.7%) of the RealStar-genus and the RealStar-species assays, respectively, were very good. The limit of detection (LoD) for the RealStar-genus assay showed a mean value of 0.28 parasites/µL with Plasmodium falciparum samples; while, the LoD of the RealStar-species assay ranged from 0.09 parasites/µL for P. vivax to two parasites/µL for P. ovale. The time to complete a diagnosis was established in 4 hours. Our findings showed a very good concordance of both assays compared with the reference method, with a very good analytical sensitivity. RealStar-species assay was able to correctly characterize double and triple infections. Therefore, these RealStar assays have shown to be useful tools in malaria diagnosis in non-endemic countries and even endemic countries, and for malaria control in general, detecting low parasitemias with sensitivity similar to the most sensitive methods as nested PCR, but with lower time to get the results.

Objective: Main malaria diagnosis is based on microscopic examination combined with rapid diagnostic tests. Both methods have low sensitivity and specificity. Loop-mediated isothermal amplification techniques have shown a sensitivity similar to PCR but with lower times of performance. This study aimed to assess a commercial LAMP for the diagnosis of malaria (Alethia^® Malaria) against the Nested-Multiplex-Malaria PCR, including the analytical sensitivity and the operational characteristics. Results: One hundred five samples out of 114 rendered valid results, obtaining 85 positive samples and 18 negative samples with an agreement of 98% compared to the reference method with a sensitivity, specificity and kappa coefficient of 98.84%, 94.74% and 0.94 respectively, with only two discrepant samples. The turnaround time was estimated in 1 h and 30 min, with a cost of 32.67€ per determination. The results showed several advantages of the Alethia^® Malaria, as it was easy to perform, minimal training requirement and 40 min run. Moreover, it includes an internal control to avoid false negatives. However, it also showed some limitations such as the need for a specific amplification and detection device, the detection of only Plasmodium spp. and a very high price.

Molecular methods are necessary to detect low-density malaria infections. The purpose of this study was to assess the diagnostic performance of six malachite-green loop-mediated amplification method (MG-LAMP) assays (MG-LAMP-Pf, MG-LAMP-Pv, MG-LAMP-Po, MG-LAMP-Pm, MG-LAMP-Pk, and MG-LAMP-Pspp) for the species-specific detection of each human Plasmodium, including P. knowlesi, and the Plasmodium genus compared with the nested-multiplex malaria polymerase chain reaction (NM-PCR), using 161 malaria-positive and 274 malaria-negative samples. MG-LAMP-Pspp assay detected the five human Plasmodium species and each species-specific MG-LAMP assay detected only its corresponding species. Sensitivity, specificity, and predictive values of MG-LAMP assays, compared with NM-PCR, were > 90%, except in the case of the MG-LAMP-Pm assay, which dropped to 47%. Limit of detection for MG-LAMP-Pspp assay ranged from 0.1 parasites/µL for P. falciparum to 16.9 parasites/µL for P. malariae samples, and it was similar for the rest of MG-LAMP assays except for the MG-LAMP-Pm assay. Turnaround time was estimated to be 2 hours and 35 minutes for one MG-LAMP assay and 4 hours and 15 minutes if all species-specific MG-LAMP is set up, whereas for the NM-PCR, turnaround time was ∼6 hours and 15 minutes. Costs per determination ranged from 1 to 6 euros for MG-LAMP assays and 5 euros for NM-PCR. Therefore, MG-LAMP assays appear to have good concordance compared with the reference method, except for the MG-LAMP-Pm assay. They can detect low parasitemia and identify malaria species, with lower costs and shorter time to obtain results, and they are suitable tools to be used in endemic and non-endemic countries for malaria detection.