Reflectance Confocal Microscopy Terminology in Spanish: A Delphi Consensus Study. / Consenso de terminología en microscopia confocal de reflectancia en español mediante método Delphi.

The terminology used to describe reflectance confocal microscopy (RCM) findings in both melanocytic and nonmelanocytic lesions has been standardized in English. We convened a panel of Spanish-speaking RCM experts and used the Delphi method to seek consensus on which Spanish terms best describe RCM findings in this setting. The experts agreed on 52 terms: 28 for melanocytic lesions and 24 for nonmelanocytic lesions. The resulting terminology will facilitate homogenization, leading to a better understanding of structures, more standardized descriptions in clinical registries, and easier interpretation of clinical reports exchanged between dermatologists.

Morphology and molecular taxonomy of the tongue worm, genus Raillietiella (Pentastomida) from the lungs of berber skinks Eumeces schneideri (Scincidae): First report.

In the present study, pentastomids belonging to the order Cephalobaenida were isolated from the lungs of Berber skinks Eumeces schneideri (Famiy: Scincidae), which were morphologically described by light and scanning electron microscopy and taxonomically justified by 18s rDNA molecular analyses of the parasites. Seventeen host specimens were collected from well-vegetated wadis at high altitudes, Jizan, Saudi Arabia as new type locality; twelve specimens (70.59%) were infected. All of the recovered parasites were adults, possessed small broadly triangular cephalothorax flattened on the ventral surface and merged smoothly with a uniformly thick and squat abdomen and terminated in a pair of divergent lobes. The results obtained indicated that the parasites belong to the sharp-tipped posterior-hook Raillietiella spp. distinguished from other raillietiedids of the same group some important characteristic features including annulus number, shape and dimensions of the buccal cadre, copulatory spicules, and anterior and posterior hooks. The anterior hook of the female specimens (n=5) had a blade length (AB) of 135±5 (110-146) µm and shank length (BC) 158±5 (150-169) µm while the posterior hook was much larger with AB measuring 221±5 (200-236) µm and BC 286±6 (280-289) µm. For the male specimens (n=5), the anterior hook had an AB of 73±3 (72-75) µm and a BC 102±5 (100-103) µm. The posterior hook was much larger with AB 190.6±5 (190-191) µm and BC 221±5 (280-289) µm. The morphological characterization of the recovered parasites was closely similar to R. aegypti previously isolated from the same host. Sequence alignment by the maximum likelihood analysis for the data obtained from the 18S rDNA analysis of the parasites exhibits identities ranging between 88-95% with pentastomid genera recovered from the GenBank. The phylogenetic tree supported the inclusion of the parasites within the monophyletic Pentastomida clade with maximum identity to the raillietiellid species. The recovered sequences from the present study were deposited in GenBank under Accession number MK970649.1. The present molecular analysis was the first to confirm the taxonomic position of R. aegypti isolated from the host examined.

LED fluorescence microscopy in the diagnosis of tuberculosis: Fading and restaining of smears for external quality assessment.

Blinded rechecking is a method proposed for external quality assurance (EQA) of auramine-stained acid-fast bacilli (AFB) smears using fluorescence microscopy (FM), however, this procedure is not well developed and slides fading over time could compromise its implementation. Since bleaching of fluorescent molecules involves temperature-dependent chemical reactions, it is likely that low temperatures could slow down this process. We stored auramine-stained slides under different environmental conditions, including -20°C, and examined them over time. The slides stored in all the environments faded. At -20°C, fading was not reduced in relation to room temperature. Restaining and re-examining smears after five months showed that the slides containing saliva and storage at -20°C were associated with failure in AFB reappearance. In conclusion, the practice of freezing slides until they are viewed should be discouraged as it has a negative effect on blinded rechecking by reducing reading concordance after restaining. Specimen quality should be considered when interpreting FM-EQA results.

Reflectance Confocal Microscopy for Inflammatory Skin Diseases.

In vivo reflectance confocal microscopy (RCM) is a relatively novel non-invasive tool for microscopic evaluation of the skin used prevalently for diagnosis and management of skin tumour. Its axial resolution, its non-invasive and easy clinical application represents the goals for a large diffusion of this technique. During the last 15 years, RCM has been demonstrated to be able to increase the sensibility and sensitivity of dermoscopy in the diagnosis of skin tumours integrating in real time clinic, dermoscopic and microscopic information useful for the definition of malignancy. Despite to date, no large comparative studies on inflammatory skin diseases has been published in the literature, several papers already showed that RCM has a potential for the evaluation of the descriptive features of the most common inflammatory skin diseases as psoriasis, lupus erythematosus, contact dermatitis and others. The aim of the application of this technique in non-neoplastic skin diseases has been prevalently focused on the possibility of clinical diagnosis confirmation, as well as therapeutic management. Moreover, the use of RCM as driver for an optimised skin biopsy has been also followed in order to reduce the number of unsuccessful histopathological examination. In this review article we describe the confocal features of the major groups of inflammatory skin disorders focusing on psoriasiform dermatitis, interface dermatitis and spongiotic dermatitis.

Diagnostic performance of fluorescent light-emitting diode microscopy for tuberculous lymphadenitis in a high-burden setting.

OBJECTIVE: Diagnosis of tuberculous lymphadenitis using fine-needle aspiration cytology is a simple and safe but low-specificity method, whereas conventional smear microscopy has variable sensitivity due to low bacterial load. We evaluated the diagnostic performance of fluorescent light-emitting diode (LED) microscopy on routinely collected fine-needle aspirates from tuberculous lymphadenitis presumptive cases. METHODS: Fine-needle aspirates were collected from patients clinically suspected of having tuberculous lymphadenitis as part of routine diagnosis. Smear preparation was performed from the aspirate and processed for cytology, conventional Ziehl-Neelsen and LED microscopy. The remaining aspirate was processed for culture on Lowenstein-Jensen media. Capilia TB-Neo test was used to differentiate M. tuberculosis complex from non-tuberculous mycobacteria. RESULT: A total of 144 tuberculous lymphadenitis presumptive cases were included. 66.7% (96/144) were positive for M. tuberculosis complex on culture. Only one isolate was identified as non-tuberculous mycobacteria. The detection rates of Ziehl-Neelsen and LED microscopy were 18.8% (27/144) and 34% (49/144), respectively. As compared to culture, sensitivity was 25.0% [95% CI: 16.3-33.7] for Ziehl-Neelsen microscopy and 45.8% [95% CI: 35.9-55.8] for LED microscopy. The specificity was 93.8% [95% CI: 86.9-100] for Ziehl-Neelsen microscopy and 89.6% [95% CI: 80.9-98.2] for LED microscopy. LED microscopy showed a statistically significant increase in sensitivity and similar specificity compared to Ziehl-Neelsen microscopy. Mean reading time of positive slides was 2.62 min/slide for Ziehl-Neelsen and 1.60 min/slide for LED microscopy. Cytology showed sensitivity of 82.3% and specificity of 54.2%. LED microscopy detected TB bacilli in 33.3% of cases cytologically classified as suppurative abscess. CONCLUSION: The LED microscopy for tuberculous lymphadenitis had significantly higher sensitivity and shorter screening time than Ziehl-Neelsen microscopy. Use of LED microscopy among cases classified as suppurative abscess on fine-needle aspirate cytology improves evidence-based diagnosis of presumptive tuberculous lymphadenitis cases. Moreover, LED microscopy could be considered as an alternative approach in settings where fine-needle aspirate cytology is impractical.

[Morphological and molecular characterization of isolates of Macrophomina phaseolina associated with sugarcane in Mexico]. / Caracterización morfológica y molecular de aislados de Macrophomina phaseolina asociados a caña de azúcar en México.

Charcoal rot caused by Macrophomina phaseolina is an important disease of sugarcane in Mexico. This study was carried out to characterize isolates of M. phaseolina obtained from sugarcane by the combination of morphological and molecular analyses. The morphological characterization of 10 isolates was performed using scanning electron microscopy and light microscopy. To confirm the morphological identification, rDNA from two representative isolates was extracted, and the internal transcribed spacer (ITS) region was amplified by polymerase chain reaction and sequenced using specific primers MpKF1 and MpKR1. Based on their morphological characteristics, all isolates were identified as M. phaseolina. Moreover, the analysis of two ITS sequences showed 100% similarity with the M. phaseolina sequences deposited in the GenBank. To our knowledge, this is the first study in the world aimed at characterizing isolates of M. phaseolina obtained from sugarcane.

Usefulness of confocal microscopy in distinguishing between basal cell carcinoma and intradermal melanocytic nevus on the face.

The clinical distinction between basal cell carcinoma (BCC) and intradermal melanocytic nevus lesions on the face can be difficult, particularly in young patients or patients with multiple nevi. Dermoscopy is a useful tool for analyzing characteristic dermoscopic features of BCC, such as cartwheel structures, maple leaf-like areas, blue-gray nests and dots, and ulceration. It also reveals arborizing telangiectatic vessels and prominent curved vessels, which are typical of BCC, and comma vessels, which are typical of intradermal melanocytic nevi. It is, however, not always easy to distinguish between these 2 conditions, even when dermoscopy is used. We describe 2 facial lesions that posed a clinical and dermoscopic challenge in two 38-year-old patients; confocal microscopy showed separation between tumor nests and stroma and polarized nuclei, which are confocal microscopy features of basal cell carcinoma.

Dermoscopic findings for the early detection of melanoma: an analysis of 200 cases.

INTRODUCTION: Dermoscopy is a complementary technique that has led to major advances in the diagnosis of pigmented skin lesions. The aim of this study was to describe the dermoscopic features of a series of melanomas and analyze the differences between melanomas in situ and invasive melanomas. MATERIAL AND METHODS: We retrospectively recorded epidemiological, clinical, histologic, and dermoscopic features of a series of 200 primary melanomas. We performed a descriptive and analytical study of the dermoscopic features identified. RESULTS: The mean age of the patients was 63 years and there was a similar distribution of male and female patients. The most common histologic subtypes were superficial spreading melanoma (62.5%) and lentigo maligna (25.5%); 67% of the melanomas had a Breslow thickness of less than 1mm and 24.5% were melanomas in situ. Overall,the most common global dermoscopic features were the multicomponent pattern (33.5%), the reticular pattern (18%), and the nonspecific pattern (15.5%). The most common local features were structureless homogeneous areas (67.5%), white-blue structures (58%), an atypical pigmented network (55.5%), and irregularly distributed dots and globules (44%). The following features were more common in invasive melanomas than in melanomas in situ: blue, gray, red and white colors, multicomponent and homogeneous patterns, dots and globules, blue-white structures, homogeneous areas, a blue-white veil, white shiny structures, a reverse pigment network, and milky-red areas. The reticular pattern was more common in melanomas in situ. DISCUSSION: The use of dermoscopy has contributed to the early diagnosis of melanoma. The most common dermoscopic features of melanoma are multiple structures and colors (multicomponent pattern), an atypical reticular pattern (with wide, irregular meshes), and an absence of distinguishing features (nonspecific pattern) associated with the presence of vascular structures. CONCLUSIONS: Dermoscopy facilitates the diagnosis of melanoma and could be useful for differentiating between melanoma in situ and invasive melanoma.

Clinical usefulness of reflectance confocal microscopy in the management of facial lentigo maligna melanoma.

Facial lentigo maligna melanoma can be a diagnostic challenge in daily clinical practice as it has similar clinical and morphological features to other lesions such as solar lentigines and pigmented actinic keratoses. Confocal microscopy is a noninvasive technique that provides real-time images of the epidermis and superficial dermis with cellular-level resolution. We describe 3 cases of suspected facial lentigo maligna that were assessed using dermoscopy and confocal microscopy before histopathology study. In the first case, diagnosed as lentigo maligna melanoma, presurgical mapping by confocal microscopy was performed to define the margins more accurately. In the second and third cases, with a clinical and dermoscopic suspicion of lentigo maligna melanoma, confocal microscopy was used to identify the optimal site for biopsy.

In vivo confocal microscopy findings of cornea and tongue mucosa in patients with Sjögren's syndrome.

INTRODUCTION AND OBJECTIVES: We aimed to investigate whether Sjögren's syndrome (SS) had distinctive findings in tongue confocal microscopy in a non-invasive manner. MATERIALS AND METHODS: This retrospective case-control study evaluated corneal and tongue confocal microscopy findings of the right eyes of 25 patients with aqueous deficient dry eye and 12 healthy volunteers without dry eye findings. There were a total of 14 patients diagnosed with SS-associated dry eye (SSDE), while 11 cases were evaluated as non-Sjögren dry eye (NSDE). RESULTS: A significant difference was observed in the dendritic cell count at the corneal subbasal nerve level between the SSDE and NSDE groups (P=.018). In SSDE group, the confocal microscopy images of dendritiform hyperreflective inflammatory cells in the tongue mucosa were in favor of inflammation. However, these findings were not found in patients with NSDE or in controls. CONCLUSIONS: This study showed that confocal microscopy provided a non-invasive evaluation of the inflammatory cells in the tongue of SS patients.

Value of multi-modality small fiber assessments in a genotypically diverse cohort of transthyretin-related amyloidosis in the early stages of disease.

INTRODUCTION: Transthyretin-related amyloidosis (ATTRv) is a progressive multisystem disorder, predominantly involving the peripheral nerve system (PNS) and heart. Quantification of small fiber damage may help guide treatment decisions, as amyloid deposits frequently affect those fibers early in disease course. Corneal confocal microscopy (CCM) is a promising method to monitor patients with ATTRv, due to similarities between corneal nerves and PNS, as the cornea is innervated by Aδ and C fibers. METHODS: We compared CCM measures from ATTRv patients to a group of healthy individuals, matched by age and gender. We then investigated the correlations between small fiber tests (SFT): CCM, LDI-Flare and CDT, COMPASS-31 and disability scales (RODS and ONLS) in patients. RESULTS: Of 20 patients (6 with V30M), mean age 50.3±15.3Y, 7 female (35%), six (30%) had polyneuropathy and 10 (50%) carpal tunnel syndrome. CDT was abnormal in 9 and LDI-flare in 6 patients. CCM was abnormal in 19 tested patients and significantly reduced when compared to controls (CNFL: 6.31±0.31 vs. 15.21±1.02mm/mm2, p<0.001). Mean COMPASS-31-scores were 22.27±22.84; RODS and ONLS were 38.15±12.33 and 2.05±2.3, with no significant differences between sub-group scores. Disease duration was significantly correlated with ONLS (0.43, p=0.05) and RODS (0.46, p=0.03). There were no significant correlations between measures of disability and SFT. CONCLUSIONS: In a diverse cohort of ATTRv patients, CCM was the most frequent abnormal measurement. CCM can be a useful test to triage patients in the early disease stages and with few or equivocal symptoms.

The association of bacterial biofilm and middle ear mucosa in patients with mucosal chronic suppurative otitis media.

OBJECTIVES: To evaluate the bacterial biofilm's role in mucosal chronic suppurative otitis media (CSOM) utilizing scanning electron microscopy (SEM). METHODS: This study involved 123 participating patients with active and inactive mucosal CSOM who underwent tympanomastoid surgery. SEM was used to examine middle ear mucosa biopsies for the development of biofilms. Middle ear discharge or mucosal swabs from patients were cultured to detect any bacterial growth. The biofilm formation was correlated to the culture results. RESULTS: The biofilm was present in 69.9 % of patients (59% of them were with active mucosal CSOM) and absent in 30.1% of the patients (70% of them were with inactive mucosal CSOM), being more statistically significant in active mucosal CSOM (p-value = 0.003). A correlation that was statistically significant was found between active mucosal CSOM and higher grades (3 and 4) of biofilms (p-value <0.05). The mucosal CSOM type and the results of the culture had a relationship that was statistically significant (p-value <0.001). 60% of patients had positive culture (70% of them were with active mucosal CSOM). There was a statistically significant relation between Pseudomonas aeruginosa bacterial growth and active mucosal CSOM (p-value = 0.004) as well as higher grades of biofilms in mucosal CSOM. CONCLUSION: Mucosal CSOM, especially the active type, is a biofilm-related disease. There is a significant relation between the state of mucosal CSOM (active or inactive) and culture results with predominance of Pseudomonas aeruginosa bacterial growth in active mucosal CSOM and in higher grades of biofilms in mucosal CSOM.

Effectiveness of endoscopy in cochlear implantation.

OBJECTIVE: Determine the effectiveness of endoscopy in cochlear implantation as compared to microscopy. METHOD: Study comparing microscopy and endoscopy in cochlear implant placement in 34 patients (23 endoscopic implants and 20 implants via microscopy), between 2014 and 2019, at the Centro Medico Naval, Mexico City. The study was performed under informed consent and according to the Council for International Organizations of Medical Sciences (CIOMS). RESULTS: Of the 34 patients, 12 were children or adolescents and 22 were adults. The visualization of the round window classified via microscopy per St. Thomas Hospital's classification showed that type IIB prevailed in 30.2% of patients, and type III in 41.9%, and when using the endoscope, the round window was observed in full in 82.6% of patients (type I), and type IIA was only observed in 17.4% (four patients). The number of attempts made to place the cochlear implant was greater with the microscope. The time to insertion of the electrode was 1.6 minutes. No differences were observed (p > 0.05) in the number of inpatient days. Cochleostomy was more frequent when using the microscope. CONCLUSIONS: Endoscopy is an effective resource in cochlear implantation for posterior tympanotomy, with no complications observed, offering greater safety in inserting the electrode through the round window.

OBJETIVO: Determinar la efectividad de la endoscopía en la implantación coclear en comparación con la técnica microscópica. MÉTODO: Se comparó la microscopía frente a la endoscopía en la colocación de implante coclear en 34 pacientes (23 endoscópicos y 20 microscópicos), del año 2014 al año 2019, en el Centro Médico Naval de la Ciudad de México. El estudio se realizó bajo consentimiento informado y apegado a las normas del Council for International Organizations of Medical Sciences. RESULTADOS: De los 34 pacientes, 12 eran niños o adolescentes y 22 eran adultos. La visualización de la ventana redonda fue clasificada con microscopio según la clasificación del St. Thomas Hospital, predominando la tipo IIB (30.2%) y la III (41.9%), y al utilizar el endoscopio se observó completa en el 82.6% (tipo I) y tipo IIA en tan solo el 17.4% (cuatro pacientes). El número de intentos en la colocación del implante coclear fue mayor con el microscopio. El tiempo en el que se insertó el electrodo fue de 1.6 minutos. No hubo diferencias (p > 0.05) en la estancia hospitalaria. Fue más frecuente la cocleostomía cuando se uso el microscopio. CONCLUSIONES: La endoscopía es un instrumento efectivo en la implantación coclear por timpanotomía posterior, sin presentarse complicaciones y dando mayor seguridad para insertar el electrodo por la ventana redonda.

Simple questionnaire and urine reagent strips compared to microscopy for the diagnosis of Schistosoma haematobium in a community in northern Ghana.

OBJECTIVES: To evaluate the utility of a simple questionnaire and urine reagent strip testing for the rapid diagnosis of Schistosoma haematobium in rural northern Ghana. METHODS: Cross-sectional parasitological and questionnaire survey in a community in northern Ghana. Participants provided two urine specimens that were examined under a microscope using a centrifugation method. The first urine sample was additionally subjected to reagent strip testing. A short questionnaire was administered to all participants. RESULTS: Microscopy of urine samples obtained from 208 individuals aged 1-77 years revealed an S. haematobium prevalence of 6.8%. The presence of any blood or protein on a urine reagent strip was 100% and 42% sensitive, and 93% and 80% specific for S. haematobium diagnosis. Questionnaires were completed by 198 individuals. Self-reported haematuria showed a sensitivity of 53% and a specificity of 85%. A dichotomous two-question panel was helpful in S. haematobium diagnosis, with working and playing near the river significantly associated with S. haematobium infection (P < 0.001). CONCLUSION: The use of urine reagent strips, coupled with questions pertaining to water contact patterns, might be considered for point-of-contact diagnosis of S. haematobium where microscopy is unavailable.

Is dark-field microscopy still useful for the primary syphilis diagnosis in the 21ST century?

INTRODUCTION: Serological test for primary syphilis could be negative the first 5-15 days. The aim of this study was to evaluate the benefit of including dark field microscopy (DFM) in the diagnosis algorythm for primary syphilis. MATERIALS/METHODS: Patients attended to a sexual transmission diseases clinic of Madrid, from 2015 to 2019, for a genital ulcer with clinical suspicion of primary syphilis. They were tested for DMF and serological test (EIA/TPPA/RPR). RESULTS: Over the total amount of samples (806), 53.2% (429) were positive for DFM. Thus, the 48% of the 429 patients had negative serological test (EIA/RPR) of which the 77.6% were positive at TPPA. CONCLUSIONS: DFM allows primary syphilis early diagnosis, even without serological test. If no direct detection methods are available, for patients without history of syphilis, TPPA could help to diagnose primary syphilis.

In vivo confocal microscopy findings after SMILE refractive surgery technique.

OBJECTIVE: To evaluate confocal microscopy finding after SMILE surgery by in vivo confocal microscopy and stromal lenticule and stromal interface accuracy planed vs measured. METHOD: Thirty eyes of 15 patients were evaluated before and 1 month after SMILE surgery by using confocal microscopy. Cellular morphology was studied. Planed stromal lenticule thickness was compared vs measured stromal lenticule thickness after surgery by comparing the total stromal corneal thickness before vs after surgery. Stromal corneal interface was evaluated and depth of this planed interface was compared vs measured post-surgery interface depth. RESULTS: Sub-epithelial nerve plexus was absent at 1 month after surgery. Activated keratocytes were found before and after stromal corneal interface depth. There was not statistical significant difference between mean planed stromal lenticule vs. post-surgery measured (102.3 ± 25.7 vs. 104.2 ± 29.9 mm; p = 0.73). There was not statistical significant difference between mean planed stromal interface depth vs. post-surgical measured (113.7 ± 8.2 vs. 120.5 ± 17.3 mm; p = 0.058). CONCLUSIONS: Confocal microscopy is useful to evaluate changes after SMILE surgery; lenticule thickness and stromal interface depth are exact.

OBJETIVO: Evaluar los hallazgos mediante microscopía confocal in vivo de pacientes operados de cirugía SMILE, la exactitud del lentículo estromal y la profundidad de la interfaz estromal planeados versus medidos. MÉTODO: Treinta ojos de 15 pacientes operados de SMILE se estudiaron mediante microscopía confocal antes y al mes de la cirugía. Se evaluaron la morfología celular, el espesor del lentículo estromal planeado versus el medido en el posoperatorio mediante la diferencia entre el espesor estromal preoperatorio y el postoperatorio, así como la interfaz estromal, y se comparó la profundidad de esta interfaz estromal planeada con la medida en el posoperatorio. RESULTADOS: Se observó la ausencia del plexo nervioso subepitelial al mes del posoperatorio y una activación de queratocitos anterior y posterior a la profundidad de la interfaz estromal. No hubo diferencia en el lentículo estromal planeado versus el medido en el posoperatorio (102.3 ± 25.7 vs. 104.2 ± 29.9 mm; p = 0.73). No hubo diferencia entre la profundidad de la interfaz estromal planeada y medida (113.7 ± 8.2 vs. 120.5 ± 17.3 mm; p = 0.058). CONCLUSIONES: Tras la cirugía SMILE se pueden evaluar los cambios mediante microscopía confocal; el espesor del lentículo y la profundidad de la interfaz son exactos.

Multi-organ pathological findings associated with COVID-19 in postmortem needle core biopsies in four patients and a review of the current literature.

Coronavirus disease 2019 (COVID-19) has spread worldwide, resulting in significant morbidity and mortality. Histopathological findings are essential in understanding its pathogenesis and we present our findings from postmortem core needle biopsies in an attempt to share information that may shed some light on this severe pandemic. Different organ samples from four patients with PCR-confirmed COVID-19 at the Infanta Sofía Hospital (Madrid) were studied during the months of April and May, 2020 by six pathologists using routine stains, histochemistry and immunohistochemistry. Results were compared with other reported cases. All patients had a clinical diagnosis of pneumonia and biopsies revealed lung damage in the majority. Heart, liver, spleen and kidney were also studied and abnormalities were found in all cases and are extensively described. The histopathology of organs affected by COVID-19 is vital to the understanding of this disease and its sequelae.

[The relevance of electron microscopy in kidney biopsies to 21st century pathology]. / Microscopía electrónica en biopsias renales: una evaluación de su utilidad en el siglo XXI.

INTRODUCTION: Electron microscopy (EM) has been used in the study of renal biopsies for more than 5 decades; however, it is expensive and the possibility of restricting it to selected cases has been considered. This study aims to reevaluate the necessity for EM in the diagnosis of renal biopsies today. MATERIAL AND METHODS: All renal biopsies taken between 2016 and 2019 with adequate light microscopy (LM), immunofluorescence (IF) and EM studies were included. The initial diagnosis (without EM) and the final diagnosis (with EM) was recorded. EM was considered necessary in cases in which the initial and final diagnoses did not concur, when diagnosis could not be made with LM and IF only or if the EM study revealed further clinically relevant findings. RESULTS: A total of 621 biopsies were included, 498 (80.2%) of native kidneys and 123 (19.8%) of transplanted kidneys. In 115 cases (18.5%) EM had been deemed necessary for diagnosis; it was required more frequently in hereditary diseases (96.8%) and isolated hematuria (88.9%) but less often in nephrotic syndrome (6.7%) and renal transplant biopsy (5.7%) (p < 0.001). CONCLUSIONS: EM was required in less than a fifth of renal biopsies, being more necessary in isolated hematuria and hereditary diseases and less so in nephrotic syndrome and in renal graft biopsies. These findings may prove useful as a guide to case selection protocols in which EM could be considered as a non-mandatory technique.

Implementation of a Gene Panel for Genetic Diagnosis of Primary Ciliary Dyskinesia. / Implementación de un panel de genes para el diagnóstico genético de la discinesia ciliar primaria.

INTRODUCTION: Primary ciliary dyskinesia (PCD) is characterized by an alteration in the ciliary structure causing difficulty in the clearance of respiratory secretions. Diagnosis is complex and based on a combination of techniques. The objective of this study was to design a gene panel including all known causative genes, and to corroborate their diagnostic utility in a cohort of Spanish patients. METHODS: This was a multicenter cross-sectional study of patients with a high suspicion of PCD, according to European Respiratory Society criteria, designed around a gene panel for massive sequencing using SeqCap EZ capture technology that included 44 genes associated with PCD. RESULTS: We included 79 patients, 53 of whom had a diagnosis of confirmed or highly probable PCD. The sensitivity of the gene panel was 81.1%, with a specificity of 100%. Candidate variants were found in some of the genes of the panel in 43 patients with PCD, 51.2% (22/43) of whom were homozygotes and 48.8% (21/43) compound heterozygotes. The most common causative genes were DNAH5 and CCDC39. We found 52 different variants, 36 of which were not previously described in the literature. CONCLUSIONS: The design and implementation of a tailored gene panel produces a high yield in the genetic diagnosis of PCD. This panel provides a better understanding of the causative factors involved in these patients and lays down the groundwork for future therapeutic approaches.