Blood Smear Slides Research Articles

Diagnostic accuracy of bone marrow blood evaluation in haemophagocytic lymphohistiocytosis paediatric patients.

Haemophagocytic lymphohistiocytosis (HLH) is a rare and serious immunological syndrome that involves a strong activation of cytotoxic T lymphocytes and macrophages. HLH determines a cytokine-mediated tissue injury with a contemporary multi-organ failure and a high fatality rate. A retrospective study was performed considering the medical records of paediatric patients who underwent a bone marrow aspirate for suspect HLH. The biomarkers evaluated were among those included in the HLH-2004. Lactate dehydrogenase (LD) was also evaluated. Haemophagocytosis was evaluated in bone marrow blood smear slides. Enrolled were 11 patients included in the HLH group and 8 patients as controls. Haemoglobin and fibrinogen resulted lower in HLH patients than in controls, while blood triglycerides, serum ferritin and LD resulted increased. Blood triglycerides and fibrinogen discriminated HLH cases perfectly, with a sensitivity and specificity of 100%. Ferritin had a sensitivity of 100% and a specificity of 83% (cut off ≥3,721µg/L) and LD of 73% and of 100% (the cut off ≥1,903U/L). Haemoglobin was found to have a sensitivity of 75% and a specificity of 100% (cut off ≤ 96g/L). Total haemophagocytes cell counts were not different between patients and controls. Only the increased number of phagocytized nucleated red blood cells (NRBC) was found to be significantly increased in the patients. Erythrocytes phagocytosis (≥4/1,000 cells) only tended towards significance. The blood biomarkers showed better diagnostic performance than the morphological evaluation. Among the different cell lineages engulfed by haemophagocytes, the best diagnostic performance was obtained by phagocytosed mature erythrocytes and immature nucleated erythrocytes.

Comparative assessment of nucleated red blood cells using a fully automated hematology analyzer versus slide examination.

Objective: To determine the sensitivity and specificity of fully automated hematology analyzer for nucleated red blood cells (NRBCs) keeping microscopic slide examination as gold standard. Study Design: Cross-sectional study. Setting: Department of Hematology, RMI Hospital, Peshawar. Period: 1st December 2023 to 29th Feb 2024. Methods: A total of 160 blood samples referred for assessment to the hematological department for the suspected abnormal NRBCs were included in the study through consecutive sampling. Fresh Blood samples were analyzed using fully automated hematology analyzer. Manual counting of the NRBCs of the same samples was done using manual blood smear slide examination. Findings of the automated nucleated red blood cell counting methods were statistically analyzed keeping microscopic slide examination as gold standard. Sensitivity, specificity, accuracy and negative and positive predictive values were calculated for diagnosis of NRBC (count >0.02 × 109/L). Clinical sensitivity of the test was determined by Receiver Operating Test (ROC). Results: The Mean±SD of patient’s age in this study was 46.78±14.89 years with an age range of 0.5 to 67 years. The male gender was 53.75% while female gender was 46.25%. Fully automated hematology analyzer detected 30 (18.75%) samples while slide examination detected 31 (19.37%) samples with NRBC count >0.02 × 109/L. Fully automated hematology analyzer has shown sensitivity of 93.55 %, specificity 99.23% and diagnostic accuracy by 98.13%. PPV was 96.67% and NPV was 98.46 (p < 0.000) for the diagnosis of NRBC. The cutoff value of 0.0175 × 109/L (20/µL) offered the best balance between sensitivity and specificity based on ROC curve. Conclusion: Association is present for the NRBC count among the fully automated hematology analyzer and microscopic slide examination.

Abstract 3768: Optimization of digital pathology through laser capture microdissection with a 405 nm laser

Abstract Tumor cellular heterogeneity is a complex problem in cancer molecular diagnostics and personalized therapy. The tumor is a product of the different types and interactions of host and immune cells. Investigators have created a variety of methods to procure separate subpopulations of the tumor microenvironment for individual analysis. One of the most successful methods of this is laser capture microdissection (LCM). This method captures specific subpopulations of cells under direct microscopic visualization. This LCM technology has successfully been used for over 20 years, revealing a variety of insights into cancer pathogenesis and mechanisms of therapeutic response employed in numerous clinical trials. Nevertheless, there are two drawbacks to the current systems. For UV cutting (365 nm), the tissue is significantly damaged by the UV light energy. Infrared (808 nm) laser systems do not damage the tissue however, this method has low power and poor resolution. Our first hypothesis to be tested is that a new class of near UV 405 nm laser will have small capture areas approaching single cells while simultaneously preventing cellular damage. The second weakness of LCM is the requirement for an open-faced tissue section without a coverslip. Moreover, when visualizing the tissue of interest, the identification of regions of interest is obscured due to the refractive index mismatch between the laser microdissection cap and the tissue surface. We hypothesize that using a volatile organic compound with a refractive index similar to glass would elucidate the tissue region of interest to allow proper visualization. We successfully developed and tested a near 405 nm wavelength LCM system that utilizes a “liquid” coverslip to visualize the sample of interest with higher clarity, resolution, and contrast for immunohistochemistry, immuno-fluorescence, blood smear slides, and conventional hematoxylin-eosin tissue staining. Overall, these critical improvements LCM technology now allow for single-cell tissue capture, system automation, tissue visualization, and opens the door for artificial intelligence-assisted spatial profiling. Citation Format: Thomas Philipson, Marissa Howard, Kevin Johnson, Amanda Still, Furkat Yunusov, Emanuel Petricoin, Virginia Espina, Noel Gonzalez, Alan Carpino, Lance Liotta. Optimization of digital pathology through laser capture microdissection with a 405 nm laser [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3768.

Abstract 3697: Proof-of-concept study to enumerate large oncosomes using the RareCyte CTC platform

Abstract Introduction: Large oncosomes (LO) are produced by tumor cells and can be detected in blood from cancer patients. The clinical utility of enumerating and characterizing LOs along with CTCs has been demonstrated, especially in settings where CTC counts are low. The RareCyte CTC platform has the potential to capture LOs along with CTCs. We performed a pilot study to evaluate the RareCyte CTC platform’s potential to analyze LOs using established preanalytical procedures. Methods: Metastatic breast cancer patients’ (N=10) blood smears labelled with RarePlex® Enumeration panel kit were re-evaluated to identify objects with LO-like characteristics at 10x magnification, along with negative control slides (N=5). LO-like objects were annotated and re-scanned at 40x magnification using the CyteFinder® imaging instrument. After establishing the criteria for LO identification, LO enumeration was performed on a cohort of MBC patients (N=6). Sets of five blood smear slides stained with the RarePlex® Enumeration panel kit were reviewed and LO counted. Total LO counts were established by extrapolating LO counts to 7.5 mL plasma. Pearson correlation was used to determine the level of correlation between LO counts and previously obtained CTC counts. Results: The established identification criteria enabled the consistent enumeration of LOs at 10X magnification. Total LO count was 9.6 on the negative control slides. In MBC patients the mean total LO count was 306.2 (range 3.2 - 792). There was a significant positive correlation between LO and CTC count (r=0.855, p=0.3). In samples with CTC>0 (N=3), the average total LOs count was 588, about six times higher than CTC count. In CTC=0 patient samples (N=3) the mean total LO count was 24.3 (range 3.2-36). Conclusions: We report a novel protocol to quantify large oncosomes using the RareCyte CTC platform using established preanalytical workflows. The strong CK/EpCam staining in LOs and the strong correlation of LO count with CTC count indicates their tumoral origin. Due to their higher abundance, LOs combined with CTCs offer a more sensitive tool for treatment response monitoring. Due to its broad range, LO counts in patients with CTC=0 may reveal therapy response dynamics undetectable with CTC enumeration alone. Ongoing work focuses on the parallel detection and integrated evaluation of HER2 by immunofluorescent staining of CTCs and LOs, along with digital PCR based HER2 quantification using cfDNA from matched blood plasma samples. Citation Format: Eszter Papp, Jo Vandesompele, Peter B. Vermeulen, Luc Y. Dirix, Mark Kockx. Proof-of-concept study to enumerate large oncosomes using the RareCyte CTC platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3697.

Evaluation of Malaria Microscopy Diagnostic Performance at 40 Public Health Facilities in Abidjan, Côte d'Ivoire in 2020.

Microscopic diagnosis of Giemsa-stained thick and thin blood films remained the gold standard laboratory method for the diagnosis of malaria. In this context, we felt it was important to conduct this evaluation with 40 public medical biology laboratories (MBLs) in the Abidjan 1 health region that perform blood parasitology tests to improve their implementation process. This descriptive and analytical study took place in July 2020 and involved participating laboratories (PLs) from the public sector in Abidjan. A set of 3 blood smear slides of variable parasite densities (PDs) with assigned values (AVs) of parasite densities and assigned Plasmodium species was used. The criterion for establishing the parasite density compliance interval was assigned values of ± 25%, and the performance rates were compared to the 80% recommended by the WHO for the African region. Nearly a quarter (11/40) of the participating laboratories had a compliance rate greater than 80%, including 10 with a performance of 100% for the ability to identify parasites. Regarding identifying plasmodial species, a concordance rate of 100% was obtained for slide 1 for Plasmodium falciparum, while this rate was 20% for slide 2 for Plasmodium ovale. For parasite densities < 200/µl, 87.5% of the participating laboratories (PLs) had a performance rate lower than 80%, while 95% of these PLs had a performance rate higher than 80% for parasitaemia > 2000/µl. There is a need to strengthen adapted to low parasitaemia, to improve the biological confirmation of malaria in Côte d'Ivoire.

Establishing Conditions for Blood Smear Drying and Staining on Sysmex SP-50 for Leukocyte Differential Count.

The purpose of this study was to determine the staining conditions and appropriate fan1 start time (FAN1ST) for Sysmex SP-50 to produce blood smears (BS) that reflect the true lymphocyte morphology of patient samples. Using different start times of fan1, we obtained a set of 84 blood smear slides from 21 blood samples and measured 10,920 lymphocyte areas, which were then converted to compare lymphocyte sizes. We also performed a leukocyte differential count using Sysmex DI-60 on 202 blood smear slides prepared before and after the change in staining conditions and compared the results. The mean lymphocyte sizes at FAN1ST 0 second, 5 seconds, 10 seconds, and 30 seconds were 12.55 μm, 12.14 μm, 11.27 μm, and 10.50 μm, respectively. The mean differences in the preclassification of neutrophils, lymphocytes, monocytes, eosinophils, and basophils in DI-60, according to the SP-50 staining conditions, were 0.88, -1.58, -0.24, 0.37, and 0.07, respectively. Wright-Giemsa staining of blood smears prepared on the SP-50 showed that changing the pH of the concentrated phosphate buffer to 6.6 and adjusting the staining time did not affect the results of the leukocyte differential count. However, since fan1 was used to dry the blood smear on the SP-50 and the lymphocyte size gradually decreased as the start time was delayed, it was necessary to set a start time for fan1 that did not affect the lymphocyte size.

Polarized Hyperspectral Microscopic Imaging for White Blood Cells on Wright-Stained Blood Smear Slides.

White blood cells, also called leukocytes, are hematopoietic cells of the immune system that are involved in protecting the body against both infectious diseases and foreign materials. The abnormal development and uncontrolled proliferation of these cells can lead to devastating cancers. Their timely recognition in the peripheral blood is critical to diagnosis and treatment. In this study, we developed a microscopic imaging system for improving the visualization of white blood cells on Wright's stained blood smear slides, with two different setups: polarized light imaging and polarized hyperspectral imaging. Based on the polarized light imaging setup, we collected the RGB images of Stokes vector parameters (S0, S1, S2, and S3) of five types of white blood cells (neutrophil, eosinophil, basophil, lymphocyte, and monocyte), and calculated the Stokes vector derived parameters: the degree of polarization (DOP), the degree of linear polarization (DOLP), and the degree of circular polarization (DOCP)). We also calculated Stokes vector data based on the polarized hyperspectral imaging setup. The preliminary results demonstrate that Stokes vector derived parameters (DOP, DOLP, and DOCP) could improve the visualization of granules in granulocytes (neutrophils, eosinophils, and basophils). Furthermore, Stokes vector derived parameters (DOP, DOLP, and DOCP) could improve the visualization of surface structures (protein patterns) of lymphocytes enabling subclassification of lymphocyte subpopulations. Finally, S2, S3, and DOCP could enhance the morphologic visualization of monocyte nucleus. We also demonstrated that the polarized hyperspectral imaging setup could provide complementary spectral information to the spatial information on different Stokes vector parameters of white blood cells. This work demonstrates that polarized light imaging & polarized hyperspectral imaging has the potential to become a strong imaging tool in the diagnosis of disorders arising from white blood cells.

A Study on Genotoxic Potential of Acephate in Clarias batrachus

Acephate is an insecticide made up of organophosphates. It is applied to food crops, citrus trees, on golf courses, in commercial or institutional buildings, and as a seed treatment. Products containing acephate can be purchased as tablets, liquids, granules, powders, and water-soluble packs. Acephate 75% brand name-Asataf insecticide manufactured by TATA RALLIS was used for the test. The solvent used was glass double distilled (g.d.d.) water. Fresh water catfish Clarias batrachus were collected from local water bodies of Cuttack district. All the fishes were acclimatized for fifteen days in laboratory aquaria containing 30L dechlorinated tap water prior to the initiation of the experiment. The peripheral blood smear slides were prepared from the blood collected by caudal incision in accordance with Al-Sabti (1986) and Das and Nanda (1986) with some modifications which were prepared animals were sacrificed after 24, 48 and 72 hours of Exposure and were used for each treatment group in both types of administrations (IP and dermal). The increased concentration of acephate directly affects our biological fish sample i.e. Clarias batrachus. Acephate is causing serious problems in fish as per our genotoxicity study of acephate on Clarias batrachus. Clarias batrachus is a commonly found fish species in fresh water habitat which includes ponds, ditches, wetlands and rice fields of India specially in Odisha.The irrational use of pesticides containing acephate in agriculture cause harmful effects on Clarias batrachus, which is a most important species of fish for maintaining the aquatic diversity.

An Intelligent Approach for Blood Cell Detection Employing Faster RCNN

A CBC (complete blood count) is an essential component of a thorough medical evaluation. The practice of medicine has had a significant impact on popular methodologies, such as manual counting and automated analyzers. The shape of blood cells and other cellular characteristics are highly sensitive to contamination levels in the human body. Tiny blood cell images are studied to spot disease and deviations from the norm that may indicate internal contamination. Correct cell segmentation enables more precise and powerful disease detection. Examining blood cells under a microscope is an important part of any pathological investigation. It emphasizes the examination of the correct malady after pinpointing its precise location and then ranking its anomalies, which plays a crucial role in determining the nature of a patient's condition, planning treatment, and evaluating the outcome of that treatment. Initially the complete blood count (CBC) test of patient is carried out if the report suggested that there is abnormality in the blood than that blood is poured on strip and blood is stained out through addition of coagulant. Expert hematologists are in short supply, particularly in underdeveloped nations, and are frequently overwhelmed. To help them with their burden, we provide a unique method for the automated assessment of family disease using artificial intelligence on blood smear images in this paper. For object detection to be useful in real life, it needs to be quick and good at finding different things in an image. There have been many improvements made to object detection, such as Convolutional Neural Network and the family of R-CNN. In our research, we employ Faster R-CNN to look for RBCs, WBCs, and Platelets in the Blood Cell Count (CBC) and Detection dataset. The number, shape, and other meta-information, or any abnormalities in the different parts of blood, could help find problems and/or diseases like leukemia, anemia, lymphoma, sickle cell disease, thrombocytopenia, leukopenia etc., early on. Our model is accurate enough to find a bounding box on the blood parts. A box is drawn along with their name around the various components of the blood smear slide.

Patterns of bone marrow infiltration in lymphomas- A retrospective descriptive analysis from a tertiary care centre in Southern India

Bone marrow biopsy (BMB) examination is the gold standard for staging of lymphoma which is interpreted along with other clinical, laboratory and radiological investigations. This study aimed to evaluate the various patterns of BMB infiltration by lymphomas which include Hodgkin and Non Hodgkin lymphoma and the usefulness of BMB compared to bone marrow aspiration (BMA). : In a period of three years, there were 212 cases which showed lymphoma infiltration in bone marrow. We assessed the peripheral blood smear (PBS), BMA and BMB slides and concordance between each was evaluated. BMB slides were assessed for the pattern of involvement. The most frequent subtype was chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) (21.70%) followed by Hodgkin lymphoma (HL)(12.73%)and follicular lymphoma (FL)(11.32%). Predominant pattern of infiltration showed by CLL/SLL cases was mixed pattern, by FL was paratrabecular pattern, mantle cell lymphoma(MCL) showed either diffuse or nodular, diffuse large B cell lymphoma (DLBCL) and Burkitt lymphoma(BL) showed predominantly diffuse pattern. Hairy cell leukemia (HCL) and peripheral T cell lymphoma (PTCL) showed an interstitial pattern. In HL, the pattern was usually focal nodular however diffuse involvement was seen in a few. Of the 212 cases, 50.94% cases had atypical lymphocytes or lymphocytosis in PBS and 81.14% cases showed infiltration in BMA. Atypical lymphoid cells may not be present at times in peripheral blood or bone marrow aspirate in spite of infiltration in the bone marrow. Different lymphomas show characteristic patterns of marrow involvement.

Enhancing the Microscopy Skills of Non-Science Majors and Nursing Microbiology Students: Promoting the Practice of Observing Multiple Fields of View Using Blood Smear Slides

Enhancing the Microscopy Skills of Non-Science Majors and Nursing Microbiology Students: Promoting the Practice of Observing Multiple Fields of View Using Blood Smear Slides

A combination of optimized threshold and deep learning-based approach to improve malaria detection and segmentation on PlasmoID dataset

Malaria is a life-threatening parasitic disease transmitted to humans by infected female Anopheles mosquitoes. Early and accurate diagnosis is crucial to reduce the high mortality rate of the disease, especially in eastern Indonesia, where limited health facilities and resources contribute to the effortless spread of the disease. In rural areas, the lack of trained parasitologists presents a significant challenge. To address this issue, a computer-aided detection (CAD) system for malaria is needed to support parasitologists in evaluating hundreds of blood smear slides every month. This study proposes a hybrid automated malaria parasite detection and segmentation method using image processing and deep learning techniques. First, an optimized double-Otsu method is proposed to generate malaria parasite patch candidates. Then, deep learning approaches are applied to recognize and segment the parasites. The proposed method is evaluated on the PlasmoID dataset, which consists of 468 malaria-infected microscopic images containing 691 malaria parasites from Indonesia. The results demonstrate that our proposed approach achieved an F1-score of 0.91 in parasite detection. Additionally, it achieved better performance in terms of sensitivity, specificity, and F1-score for parasite segmentation compared to original semantic segmentation methods. These findings highlight the potential of this study to be implemented in CAD malaria detection, which could significantly improve malaria diagnosis in resource-limited areas.

A Deep Transfer Learning-Based Comparative Study for Detection of Malaria Disease

Malaria is a disease caused by a parasite. The parasite is transmitted to humans through the bite of infected mosquitoes. Thousands of people die every year due to malaria. When this disease is diagnosed early, it can be fully treated with medication. Diagnosis of malaria can be made according to the presence of parasites in the blood taken from the patient. In this study, malaria detection and diagnosis study were performed using The Malaria dataset containing a total of 27,558 cell images with samples of equally parasitized and uninfected cells from thin blood smear slide images of segmented cells. It is possible to detect malaria from microscopic blood smear images via modern deep learning techniques. In this study, 5 of the popular convolutional neural network architectures for malaria detection from cell images were retrained to find the best combination of architecture and learning algorithm. AlexNet, GoogLeNet, ResNet-50, MobileNet-v2, VGG-16 architectures from pre-trained networks were used, their hyperparameters were adjusted and their performances were compared. In this study, a maximum 96.53% accuracy rate was achieved with MobileNet-v2 architecture using the adam learning algorithm

Performance of digital morphology analyzer CellaVision DC-1.

CellaVision DC-1 (DC-1, Sysmex, Kobe, Japan) is a newly launched digital morphology analyzer that was developed mainly for small to medium-volume laboratories. We evaluated the precision, qualitative performance, comparison of cell counts between DC-1 and manual counting, and turnaround time (TAT) of DC-1. Using five peripheral blood smear (PBS) slides spanning normal white blood cell (WBC) range, precision and qualitative performance of DC-1 were evaluated according to the Clinical and Laboratory Standards Institute (CLSI) EP15-A3, EP15-Ed3-IG1, and EP12-A2 guidelines. Cell counts of DC-1 and manual counting were compared according to the CLSI EP 09C-ED3 guidelines, and TAT of DC-1 was also compared with TAT of manual counting. DC-1 showed excellent precision (%CV, 0.0-3.5%), high specificity (98.9-100.0%), and high negative predictive value (98.4-100.0%) in 18cell classes (12 WBC classes and six non-WBC classes). However, DC-1 showed 0% of positive predictive value in seven cell classes (metamyelocytes, myelocytes, promyelocytes, blasts, plasma cells, nucleated red blood cells, and unidentified). The largest absolute mean differences (%) of DC-1 vs. manual counting was 2.74. Total TAT (min:s) was comparable between DC-1 (8:55) and manual counting (8:55). This is the first study that comprehensively evaluated the performance of DC-1 including its TAT. DC-1 has a reliable performance that can be used in small to medium-volume laboratories for assisting PBS review. However, DC-1 may make unnecessary workload for cell verification in some cell classes.

A telepathology based screening tool for COVID-19 by leveraging morphological changes related to leukocytes in peripheral blood smears.

As we approach the aftermath of a global pandemic caused by Severe Acute Respiratory Syndrome-Corona Virus (SARS-CoV-2), the importance of quickly developing rapid screening tests has become very clear from the point of view of containment and also saving lives. Here, we present an explorative study to develop a telepathology-based screening tool using peripheral blood smears (PBS) to identify Coronavirus Disease (COVID-19)-positive cases from a group of 138 patients with flu-like symptoms, consisting of 82 positive and 56 negative samples. Stained blood smear slides were imaged using an automated slide scanner (AI 100) and the images uploaded to the cloud were analyzed by a pathologist to generate semi-quantitative leukocyte morphology-related data. These telepathology data were compared with the data generated from manual microscopy of the same set of smear slides and also the same pathologist. Besides good correlation between the data from telepathology and manual microscopy, we were able to achieve a sensitivity and specificity of 0.83 and 0.71, respectively, for identifying positive and negative COVID-19 cases using a six-parameter combination associated with leukocyte morphology. The morphological features included plasmacytoid cells, neutrophil dysplastic promyelocyte, neutrophil blast-like cells, apoptotic cells, smudged neutrophil, and neutrophil-to-immature granulocyte ratio. Although Polymerase Chain Reaction (PCR) and antibody tests have a superior performance, the PBS-based telepathology tool presented here has the potential to be an interim screening tool in resource-limited settings in underdeveloped and developing countries.

Atypical imported loiasis characterised by recurrent anaemia in a patient from China

In March, 2021, a 49-year-old man was admitted to The Fourth People's Hospital of Nanning (Nanning, China) with recurrent dizziness and fatigue that had persisted for more than 1 year. Physical examination revealed paleness on his face and a slightly yellow sclera. He had no hepatomegaly, but he had splenomegaly. His laboratory results showed a low haemoglobin concentration of 84 g/L, a normal blood eosinophil percentage of 3·1%, a high reticulocyte percentage of 4·6%, haemoglobinuria (score of 2+), a low haptoglobin concentration of 1·01 mg/dL (normal range 32–205 mg/dL), a high lactic acid dehydrogenase concentration of 750 U/L (normal range 120–160 U/L), and a high 3-hydroxybutyrate dehydrogenase concentration of 711 U/L (normal range 72–182 U/L). He had a history of haemolytic anaemia, and the lowest haemoglobin concentration recorded was 46 g/L. The patient had previously received four blood transfusions. He had a history of malaria in Ghana and the Democratic Republic of the Congo and thus was tested for malaria. However, microfilariae (an average of 180 per mL) were incidentally discovered on examining peripheral blood samples collected on the day and night of admission. The microfilariae were 225–251 μm long and 6–8 μm wide (figure). A bone marrow smear showed two microfilariae; however, cerebrospinal fluid and urine samples showed no microfilariae. PCR identified a 286 bp filarial-specific internal transcribed spacer 1 (ITS1) fragment and a 712 bp fragment of mitochondrial cytochrome c oxidase subunit 1 (COX1). Analysis with the basic local alignment search tool on the National Center for Biotechnology Information website showed that the ITS1 and COX1 fragments had the highest similarity with those of Loa loa (99% [MK978707] and 95% [HQ186250], respectively). Review of the patient's malaria thick blood smear slide from March, 2020, revealed the presence of two microfilariae, which has not been reported previously. Additionally, in June, 2020, liver fluke eggs in the patient's stool had been reported previously and he had received albendazole (650 mg) twice daily for 7 days. Following the identification of L loa, a diagnosis of atypical imported loiasis was established and the patient was treated at our hospital with a single dose of ivermectin (150 μg/kg) orally for anti-filariasis treatment, without intervention for anaemia. 2 weeks after treatment, a peripheral blood smear showed that the number of microfilariae had decreased to nine per mL on average, and his haemoglobin concentration had increased to 120 g/L and his eosinophil percentage had increased to 22·1%. 6 weeks after treatment, his haemoglobin concentration had increased to 147 g/L, with no microfilariae detected in his peripheral blood, suggesting that his anaemia was caused by filariasis.

Malaria Parasite Detection using Efficient Neural Ensembles

Caused by the bite of the Anopheles mosquito infected with the parasite of genus Plasmodium, malaria has remained a major burden towards healthcare for years with an approximate 400,000 deaths reported globally every year. The traditional diagnosis process for malaria involves an examination of the blood smear slide under the microscope. This process is not only time consuming but also requires pathologists to be highly skilled in their work. Timely diagnosis and availability of robust diagnostic facilities and skilled laboratory technicians are very much vital to reduce the mortality rate. This study aims to build a robust system by applying deep learning techniques such as transfer learning and snapshot ensembling to automate the detection of the parasite in the thin blood smear images. All the models were evaluated against the following metrics - F1 score, Accuracy, Precision, Recall, Mathews Correlation Coefficient (MCC), Area Under the Receiver Operating Characteristics (AUC-ROC) and the Area under the Precision Recall curve (AUC-PR). The snapshot ensembling model created by combining the snapshots of the EfficientNet-B0 pre-trained model outperformed every other model achieving a f1 score - 99.37%, precision - 99.52% and recall - 99.23%. The results show the potential of model ensembles which combine the predictive power of multiple weal models to create a single efficient model that is better equipped to handle the real world data. The GradCAM experiment displayed the gradient activation maps of the last convolution layer to visually explicate where and what a model sees in an image to classify them into a particular class. The models in this study correctly activate the stained parasitic region of interest in the thin blood smear images. Such visuals make the model more transparent, explainable, and trustworthy which are very much essential for deploying AI based models in the healthcare network.

Malaria Parasite Detection using Efficient Neural Ensembles

Caused by the bite of the Anopheles mosquito infected with the parasite of genus Plasmodium, malaria has remained a major burden towards healthcare for years with an approximate 400,000 deaths reported globally every year. The traditional diagnosis process for malaria involves an examination of the blood smear slide under the microscope. This process is not only time consuming but also requires pathologists to be highly skilled in their work. Timely diagnosis and availability of robust diagnostic facilities and skilled laboratory technicians are very much vital to reduce the mortality rate. This study aims to build a robust system by applying deep learning techniques such as transfer learning and snapshot ensembling to automate the detection of the parasite in the thin blood smear images. All the models were evaluated against the following metrics - F1 score, Accuracy, Precision, Recall, Mathews Correlation Coefficient (MCC), Area Under the Receiver Operating Characteristics (AUC-ROC) and the Area under the Precision Recall curve (AUC-PR). The snapshot ensembling model created by combining the snapshots of the EfficientNet-B0 pre-trained model outperformed every other model achieving a f1 score - 99.37%, precision - 99.52% and recall - 99.23%. The results show the potential of model ensembles which combine the predictive power of multiple weal models to create a single efficient model that is better equipped to handle the real world data. The GradCAM experiment displayed the gradient activation maps of the last convolution layer to visually explicate where and what a model sees in an image to classify them into a particular class. The models in this study correctly activate the stained parasitic region of interest in the thin blood smear images. Such visuals make the model more transparent, explainable, and trustworthy which are very much essential for deploying AI based models in the healthcare network.

Évaluation Externe de la Qualité de la Goutte Épaisse/Frottis Sanguin pour le Diagnostic du Paludisme dans les Districts Sanitaires de Lomé et du Golfe au Togo

RésuméObjectifsCette étude a pour but d'évaluer la performance de la goutte épaisse/frottis sanguin aux fins d'amélioration de son processus de réalisation.Matriel et MthodesIl s'est agi d'une étude transversale descriptive réalisée de mai à juin 2017 et portant sur des laboratoires relevant des secteurs public, libéral et confessionnel à Lomé, évalués sur le diagnostic parasitologique du paludisme. Une série de 13 lames de goutte épaisse/frottis sanguin de densités parasitaires variables avec des valeurs assignées (VA) de densité parasitaire et l'espèce plasmodiale affectées, a été utilisée. Le critère de conformité de la densité parasitaire était VA ± 25% et les taux de performance ont été comparés aux 80% recommandés par l'OMS-AFRO.Résultats41,9% (13/31) des laboratoires participants avaient un taux de conformité supérieur à 80% dont 4 avec une performance de 100% pour l'aptitude à déterminer l'espèce plasmodiale. Pour les parasitémies < 100/μl, 51,6% des laboratoires participants avaient un taux de performance inférieur à 80% alors que 100% de ces laboratoires avaient un taux de performance supérieur à 80% pour les parasitémies > 2000/μl.ConclusionLes laboratoires évalués avaient des capacités insuffisantes pour l'identification de Plasmodium falciparum et l'estimation correcte des faibles parasitémies. Une nécessité de renforcement d'aptitudes techniques, adaptées au contexte de faible parasitémie s'impose pour améliorer la confirmation biologique du paludisme au Togo.

Prevalence of Oxyuriasis and its Relationship With Eosinophilia in Children Aged 1 to 6 Years in Marand City, East Azerbaijan Province, Iran

Introduction: Enterobius vermicularis, commonly known as pinworm, is one of the most prevalent intestinal nematodes of humankind belonging to the Oxyuridae family. It is a small, white or transparent, threadlike roundworm. The length of the male worm is 2 to 3 mm and the female worm is 3 to 8 mm in length. This study was performed to determine the prevalence of E. vermicularis and its relationship with disease symptoms, allergic disease symptoms, and eosinophilia in children aged one to six years in Marand city of East Azerbaijan province, Iran. Methods: In this descriptive-analytical study, 200 children were studied using systematic random sampling. Data collection method was based on a questionnaire, peripheral blood smear, and Scotch tape test. In the designed questionnaire, children’s characteristics such as gender and age and parents’ characteristics including job and level of education were recorded. Peripheral blood smear slides were obtained from each child and examined for eosinophilia by a hematologist. Scotch test samples were prepared by parents after providing the necessary training on its sampling method and then examined in the laboratory for the presence of E. vermicularis eggs by a parasitologist. Results: In this study, 64 (32%) children were diagnosed with oxyuriasis. The prevalence of oxyuriasis was 36.5% in boys and 43.7% in girls. There was no significant relationship between oxyuriasis and its symptoms. Additionally, there was no significant relationship between oxyuriasis and allergic disease symptoms. Fourteen children had eosinophilia (7%), of which 9 (64%) were infected with E. vermicularis. There was a significant relationship between E. vermicularis infection and eosinophilia (P ˂ 0.05). Conclusion: According to the results of the study and the importance of the issue for the control and prevention of this disease, it is suggested that people should be provided with the necessary information about the life cycle, ways of transmission, and especially ways to prevent E. vermicularis infestation, and basic measures should be taken to improve the economic situation of the people of the region and to promote public and personal health through public education. Finally, medical treatment of patients and their families should be considered at the same time.