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Objectives: Loa loa and Mansonella perstans are two very common filarial species in Africa. Although microscopy is the traditional diagnostic method for human filariasis, several polymerase chain reaction (PCR) methods have emerged as an alternative approach for identifying filarial parasites. The aim of this study is to compare three molecular methods and decide which is the most suitable for diagnosing human loiasis and mansonellosis in non-endemic regions using dried blood spot (DBS) as a medium for sample collection and storage., Methods: A total of 100 DBS samples, with their corresponding thin and thick blood smears, were selected for this study. Microscopy was used as the reference method to diagnose and calculate the microfilaraemia. Filarial DNA was extracted using the saponin/Chelex method and the DNA isolated was assayed by Filaria-real time-PCR, filaria-nested PCR, and cytochrome oxidase I PCR. All PCR products were subsequently purified and sequenced. The statistical values for each molecular test were calculated and compared., Results: Overall, 64 samples were identified as negative by all tests and a further 36 samples were positive by at least one of the methods used. The sensitivity and specificity were similar for the different molecular methods, all of which demonstrated good agreement with microscopy., Conclusions: Based on this study, and from a practical point of view (single and short amplification round), the optimal technique for diagnosing filarial infection in non-endemic regions is filaria-real time-PCR, which presents high sensitivity and specificity and is also able to detect a wide range of human filariae., (© 2022 John Wiley & Sons Ltd.)

Aims: Tumour-infiltrating lymphocytes have prognostic value in malignant melanoma. High endothelial venules (HEVs) are specialized vessels present in lymph nodes and tertiary lymphoid organs. CCL19, CCL21 and CCR7 regulate lymphocyte migration through HEVs. The aim of our study was to correlate HEV density in cutaneous primary and metastatic malignant melanomas with clinicopathological parameters, and with CCL19, CCL21 and CCR7 mRNA expression., Methods and Results: High endothelial venule density was evaluated by immunohistochemistry with a specific antibody, MECA-79, and chemokine expression was evaluated by real-time PCR. MECA-79-positive vessels, covered by cuboidal (C-HEV) or flat (F-HEV) endothelium, were detected in 55% of melanomas. HEV density was higher in primary melanomas than in metastases. Positive correlations were found between C-HEV density and lymphocytic infiltration, and between F-HEV density and tumour regression. Cases in which the number of C-HEVs exceeded that of F-HEVs had higher levels of CCL19, CCL21, and CCR7., Conclusions: Our results support a predominant role for C-HEV in the recruitment of lymphocytes in cutaneous melanomas, mediated by CCL19 and CCL21, whereas the density of F-HEV strongly correlates with tumour regression, Therefore, cuboidal and flat HEVs may serve as indicators of the active and late quiescent phases, respectively, of tumour regression in cutaneous malignant melanoma., (© 2013 John Wiley & Sons Ltd.)