O-237 A non-invasive approach for aneuploidy analysis in clinical miscarriages

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
N. Balaguer Cuenca ◽  
L Rodrigo ◽  
E Mateu-Brull ◽  
I Campos-Galindo ◽  
N Al-Asmar ◽  
...  

Abstract Study question Is maternal cell-free DNA (cfDNA) testing a feasible alternative to the analysis of the product of conception (POC) in clinical miscarriages? Summary answer This study demonstrates that genome-wide cfDNA testing in the maternal bloodstream constitutes a reliable tool to analyse chromosome aneuploidies in clinical miscarriages. What is known already It is well established that 50-70% of clinical miscarriages are caused by numerical chromosomal anomalies (aneuploidies), mostly trisomies. To date, conventional cytogenetic and advanced molecular techniques are used for the analysis of POC to identify the genetic cause of miscarriage, providing valuable information for genetic counselling. However, both approaches are based in the direct analysis of the abortive tissue, which entails several limitations due to the risk of culture failure and/or maternal cell contamination. To solve these drawbacks, maternal cfDNA testing emerges as a promising alternative due to the accumulated evidence. Study design, size, duration This was a retrospective study conducted in a reference genetic laboratory from January to December 2020. Before carrying out the foetal tissues collection that precludes the POC analysis, a blood sample was drawn to evaluate possible aneuploidies by cfDNA testing. Using NGS+STR POC results as the gold standard, results derived from both studies were compared to assess the percentage of concordance and the cases of non-informativeness (foetal fraction (FF) <2%), false positives, and false negatives. Participants/materials, setting, methods A total of 12 cases were included in the study. cfDNA testing in the mother’s blood was performed by using Illumina’s technology platform. Genetic testing for POC was done using an NGS technology (Thermo Fisher Scientific, USA) for 24 chromosome aneuploidy screening. Short-tandem repeat (STR) analysis allowed us to detect or rule out maternal cell contamination (MCC) and some types of polyploidies. Main results and the role of chance The non-informative rate for both analysis techniques was 9.1% (1 out of 12 cases: 1 low FF for cfDNA testing and maternal cell contamination for POC analysis). The median cfDNA FF was 9.0%. Using the molecular POC analysis as gold standard, the concordance rate between both studies was 90.0% (9 out of 10 cases;1 monosomy X, 1 trisomy (T) 21, 1 T22, 1 T11 and 5 patients with no alteration detected). No mosaics or structural rearrangements were identified by either of the two analysis techniques. The only discordant result was a case in which cytogenetics of POCs identified a triploidy. This discordancy is expected since triploidies are outside the scope of cfDNA testing. Also, foetal sex was correctly assigned in all informative cases. The sensibility and specificity of the study were estimated at 80.0 (4/5) and 100.0% (6/6), respectively. Statistics analysis suggested that no significant difference was found between both techniques regarding the aneuploidy detection ability (P=0.5). These promising results indicate that genome-wide cfDNA-based screening provides a non-invasive approach for determining whether foetal aneuploidy could explain the loss in patients experiencing early o recurrent pregnancy loss (RPL). Limitations, reasons for caution The sample size prevents drawing more significant conclusions regarding the diagnosis power similarity between both testing techniques. Therefore, a larger cohort will be essential to improve confirm the cfDNA testing performance. Current cfDNA testing technology fails in polyploidy identification, which is a potential cause of pregnancy loss. Wider implications of the findings CfDNA testing could be an alternative to POC analysis in clinical miscarriage. If optimized, cfDNA testing could be used contingently with the molecular POC analysis in cases where maternal cell contamination is present. As a result, the overall success rate in the POC program could be substantially improved. Trial registration number NA

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Brian C.-H. Chiu ◽  
Chang Chen ◽  
Qiancheng You ◽  
Rudyard Chiu ◽  
Girish Venkataraman ◽  
...  

AbstractThe 5-methylcytosines (5mC) have been implicated in the pathogenesis of diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL). However, the role of 5-hydroxymethylcytosines (5hmC) that are generated from 5mC through active demethylation, in lymphomagenesis is unknown. We profiled genome-wide 5hmC in circulating cell-free DNA (cfDNA) from 73 newly diagnosed patients with DLBCL and FL. We identified 294 differentially modified genes between DLBCL and FL. The differential 5hmC in the DLBCL/FL-differentiating genes co-localized with enhancer marks H3K4me1 and H3K27ac. A four-gene panel (CNN2, HMG20B, ACRBP, IZUMO1) robustly represented the overall 5hmC modification pattern that distinguished FL from DLBCL with an area under curve of 88.5% in the testing set. The median 5hmC modification levels in signature genes showed potential for separating patients for risk of all-cause mortality. This study provides evidence that genome-wide 5hmC profiles in cfDNA differ between DLBCL and FL and could be exploited as a non-invasive approach.


2020 ◽  
pp. jclinpath-2020-206834 ◽  
Author(s):  
Dipak Sapkota ◽  
Tine Merete Søland ◽  
Hilde Kanli Galtung ◽  
Lars Peter Sand ◽  
Simone Giannecchini ◽  
...  

The COVID-19 (caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)) epidemic started in Wuhan (Hubei Province, China) in mid-December 2019 and quickly spread across the world as a pandemic. As a key to tracing the disease and to implement strategies aimed at breaking the chain of disease transmission, extensive testing for SARS-CoV-2 was suggested. Although nasopharyngeal/oropharyngeal swabs are the most commonly used biological samples for SARS-CoV-2 diagnosis, they have a number of limitations related to sample collection and healthcare personnel safety. In this context, saliva is emerging as a promising alternative to nasopharyngeal/oropharyngeal swabs for COVID-19 diagnosis and monitoring. Saliva collection, being a non-invasive approach with possibility for self-collection, circumvents to a great extent the limitations associated with the use of nasopharyngeal/oropharyngeal swabs. In addition, various salivary biomarkers including the salivary metabolomics offer a high promise to be useful for better understanding of COVID-19 and possibly in the identification of patients with various degrees of severity, including asymptomatic carriers. This review summarises the clinical and scientific basis for the potential use of saliva for COVID-19 diagnosis and disease monitoring. Additionally, we discuss saliva-based biomarkers and their potential clinical and research applications related to COVID-19.


Author(s):  
Michael O’Brien ◽  
Yifei Dai ◽  
Glen L. Niebur

The New Zealand White (NZW) rabbit is often used as an animal model in posterolateral intrertransverse lumbar fusions [1]. Qualitative measures, such as manual palpation and radiographs, have been used to assess fusion mass [1]. However, an evaluation by sacrifice and biomechanical testing is the gold standard for quantitative data. A non-invasive approach to quantitatively assess the mechanics of spinal fusion could allow longitudinal studies with decreased need for animal sacrifice.


Gut ◽  
2019 ◽  
Vol 68 (12) ◽  
pp. 2195-2205 ◽  
Author(s):  
Jiabin Cai ◽  
Lei Chen ◽  
Zhou Zhang ◽  
Xinyu Zhang ◽  
Xingyu Lu ◽  
...  

ObjectiveThe lack of highly sensitive and specific diagnostic biomarkers is a major contributor to the poor outcomes of patients with hepatocellular carcinoma (HCC). We sought to develop a non-invasive diagnostic approach using circulating cell-free DNA (cfDNA) for the early detection of HCC.DesignApplying the 5hmC-Seal technique, we obtained genome-wide 5-hydroxymethylcytosines (5hmC) in cfDNA samples from 2554 Chinese subjects: 1204 patients with HCC, 392 patients with chronic hepatitis B virus infection (CHB) or liver cirrhosis (LC) and 958 healthy individuals and patients with benign liver lesions. A diagnostic model for early HCC was developed through case-control analyses using the elastic net regularisation for feature selection.ResultsThe 5hmC-Seal data from patients with HCC showed a genome-wide distribution enriched with liver-derived enhancer marks. We developed a 32-gene diagnostic model that accurately distinguished early HCC (stage 0/A) based on the Barcelona Clinic Liver Cancer staging system from non-HCC (validation set: area under curve (AUC)=88.4%; (95% CI 85.8% to 91.1%)), showing superior performance over α-fetoprotein (AFP). Besides detecting patients with early stage or small tumours (eg, ≤2.0 cm) from non-HCC, the 5hmC model showed high capacity for distinguishing early HCC from high risk subjects with CHB or LC history (validation set: AUC=84.6%; (95% CI 80.6% to 88.7%)), also significantly outperforming AFP. Furthermore, the 5hmC diagnostic model appeared to be independent from potential confounders (eg, smoking/alcohol intake history).ConclusionWe have developed and validated a non-invasive approach with clinical application potential for the early detection of HCC that are still surgically resectable in high risk individuals.


2021 ◽  
Author(s):  
Mina Tutunfroush ◽  
Saeid Ghorbian ◽  
Jafar Mohseni ◽  
Shahla Danaii

Abstract Recently, circulating microRNAs have attracted much attention because they can serve as reliable non-invasive diagnostic and prognostic biomarkers for pregnancy-related complications. So, this study aimed to quantify miR-23a-3p, miR-101-3p and miR-let-7c expression levels in plasma of patients with idiopathic recurrent pregnancy loss (iRPL) and healthy subjects and to evaluate their potential diagnostic value in iRPL patients. A total of 120 plasma samples were obtained from sixty women with a history of at least two consecutive iRPL and sixty healthy women without a history of miscarriage to evaluate the expression levels of the circulating miR-23a-3p, miR-101-3p and miR-let-7c by quantitative real-time polymerase chain reaction (qPCR) technique. The correlation between studied miRNAs and clinicopathological parameters was also assessed. Receiver operating characteristic (ROC) curve was plotted to determine the diagnostic accuracy of miR-23a-3p, miR-101-3p and miR-let-7c in iRPL. Our results showed that the miR-23a-3p expression level in plasma of iRPL patients was lower than those in healthy controls but without a statistically significant difference (P = 0.113). The expression levels of miR-101-3p and miR-let-7c were significantly downregulated in iRPL patients compared with healthy subjects (P < 0.05). The expression levels of miR-23a-3p and miR-let-7c was negatively correlated with number of abortions in iRPL patients. We observed statistically significant positive correlation between miR-23a-3p and miR-101-3p (r = 0.478, P = 0.001), miR-23a-3p and miR-let-7c (r = 0.561, P = 0.0001), miR-101-3p and miR-let-7c (r = 0.533, P = 0.0001) in patients with iRPL. The current study provides evidence indicating that downregulation of miR-23a-3p, miR-101-3p and miR-let-7c may be associated with iRPL.


2020 ◽  
Vol 14 (4) ◽  
pp. 7600-7608
Author(s):  
W. A. A Saad ◽  
Mohd Azuwan Mat Dzahir ◽  
Yamamoto Shinichirou ◽  
Mohamed Hussein ◽  
Maziah Mohamad ◽  
...  

The change of the spinal curvature in completing a variety of daily tasks is essential to independent living. There is still a lack of studies highlighting the lumbar segmental contribution during sit-to-stand (STS) and stand-to-flexion (STF) using non-invasive study. The purpose of this study is to compare the spine kinematics by defining lumbar as a single and multi-segmental during continuous daily motion in healthy Asian adults using a non-invasive approach. During STS, most subjects implemented kyphotic lumbar curve during the early stage of motion which revealed poor posture implementation and significant differences in the lumbar kinematics which were only noticeable at specific phases between both approaches. A significant difference in multi-segmental behaviour was observed only at the end of the motion. All three segments displayed different time responses during the transition from kyphotic to lordotic curve. Passive/delayed behavior within the lower lumbar segment was observed between 0-50% of motion completion. During STF, statistically significant differences were found between assuming lumbar as a single and multi-segment in all phases. This in vitro study identified characteristic motion patterns in the lumbar spine during daily motions. The results provided a clear description of the healthy spinal condition of adults and may serve to identify specific multi-segmental contribution.


Author(s):  
Karen Araújo Magalhães ◽  
Kamily Fagundes Pussi ◽  
Hélton Krisman de Araújo ◽  
Silvia Barbosa do Carmo ◽  
Elisabete Friozi ◽  
...  

Abstract The dog is the main domestic reservoir of Leishmania and font of infection for the vector, constituting an important host for the transmission of the parasite to humans. Non-invasive collection of swab samples for leishmaniasis diagnosis has been a promising alternative. This study analyzed the positivity of polymerase chain reaction (PCR) for the diagnosis of canine leishmaniasis in conjunctiva samples. DNA extraction was performed using SDS 20% and PCR was performed using 13A/13B primers that amplify 120-bp of Leishmania kDNA. Of the 77 dogs analyzed, 50 (64.93%) had ocular changes: 25 (32.47%) dogs had periocular lesion, 41 (53.25%) dogs had purulent eye discharge, and 17 (22.08%) dogs had both signals. PCR was positive in 35 dogs (45.45%), and there was no significant difference between dogs with and without ocular signals (p=0.4074). PCR positivity was significant higher in dogs without periocular injury (p=0.0018). Conjunctive PCR, a less invasive, fast, and painless collection technique, is indicated to complement the diagnosis, especially in dogs without periocular injury, independent of the presence of purulent eye discharge.


2015 ◽  
Vol 21 (4) ◽  
pp. 490-498 ◽  
Author(s):  
Sabine Ott ◽  
Philipp Gölitz ◽  
Edyta Adamek ◽  
Kevin Royalty ◽  
Arnd Doerfler ◽  
...  

We compared flat-detector computed tomography angiography (FD-CTA) to multislice computed tomography (MS-CTA) and digital subtracted angiography (DSA) for the visualization of experimental aneurysms treated with stents, coils or a combination of both. In 20 rabbits, aneurysms were created using the rabbit elastase aneurysm model. Seven aneurysms were treated with coils, seven with coils and stents, and six with self-expandable stents alone. Imaging was performed by DSA, MS-CTA and FD-CTA immediately after treatment. Multiplanar reconstruction (MPR) was performed and two experienced reviewers compared aneurysm/coil package size, aneurysm occlusion, stent diameters and artifacts for each modality. In aneurysms treated with stents alone, the visualization of the aneurysms was identical in all three imaging modalities. Residual aneurysm perfusion was present in two cases and visible in DSA and FD-CTA but not in MS-CTA. The diameter of coil-packages was overestimated in MS-CT by 56% and only by 16% in FD-CTA compared to DSA ( p < 0.05). The diameter of stents was identical for DSA and FD-CTA and was significantly overestimated in MS-CTA ( p < 0.05). Beam/metal hardening artifacts impaired image quality more severely in MS-CTA compared to FD-CTA. MS-CTA is impaired by blooming and beam/metal hardening artifacts in the visualization of implanted devices. There was no significant difference between measurements made with noninvasive FD-CTA compared to gold standard of DSA after stenting and after coiling/stent-assisted coiling of aneurysms. FD-CTA may be considered as a non-invasive alternative to the gold standard 2D DSA in selected patients that require follow up imaging after stenting.


2020 ◽  
Vol 73 (4) ◽  
pp. 148-152
Author(s):  
Kornél Vajda ◽  
László Sikorszki

Összefoglaló. Bevezetés: A laparoszkópia térhódítása a jobb oldali colon műtéteknél is nyilvánvaló. Ma legtöbb helyen a laparoszkóposan asszisztált jobb oldali hemikolektómia extrakorporális anasztomózissal a gold standard. A morbiditás randomizált vizsgálatok alapján még 30% körüli. A technikai fejlődés lehetővé tette az intrakorporális anasztomózist. Célkitűzés: Retrospektív módon elemezni rosszindulatú jobb oldali vastagbéldaganat miatt végzett laparoszkópos hemikolektómiák rövid távú eredményeit a két módszer összehasonlításával. Eredmények: 2018. 01. 01. – 2019. 12. 31. között 184 jobb oldali hemikolektómiát végeztünk, ezek közül 122 történt malignus betegség miatt. 51 esetben nyitott és 71 esetben laparoszkópos műtét történt. 37 férfi (átlagéletkor: 70,59 év) és 34 nő (átlagéletkor: 72,14 év) volt. 50 esetben extrakorporális (EA) és 21 esetben pedig intrakorporális anasztomózist (IA) végeztünk. Az EA csoportban 18, míg az IA csoportban 3 szövődmény alakult ki 30 napon belül (p = 0,067). Az EA csoportból 3, az IA csoportból 1 beteget veszítettünk el 30 napon belül (p = 0,66). Az átlagos ápolási idő az EA csoportban 9,48 (5–32) nap, míg az IA csoportban 6,52 (4–19) nap volt (p = 0,001) a szövődményes esetekkel együtt. A szövődményes esetek nélkül az EA csoportban 6,35 (5–10) nap, az IA csoportban pedig 5,55 (4–8) napnak bizonyult (p = 0,09). A műtéti idő pedig az EA csoportban 147 (90–240) perc, az IA csoportban pedig 146,47 (90–265) perc volt (p = 0,11). Konklúzió: Az irodalommal összhangban azt találtuk, hogy IA esetén kevesebb a szövődmény, ezzel is összefüggésben rövidebb az átlagos ápolási idő, és a műtéti időt tekintve nincs szignifikáns különbség. Ezeket figyelembe véve az intrakorporális anasztomózis javasolható jobb oldali laparoszkópos hemikolektómia esetén. Summary. Introduction: Laparoscopy became evident for right-sided colon surgery too. Today the laparoscopic-assisted right-hemicolectomy is the gold standard with extracorporeal anastomosis. Morbidity according to randomized trials is still approximately 30%. The development of the surgical technique resulted in the creation of intracorporeal anastomosis. Our aim was to compare the short-term results of the two methods. Aim: To analyse the short-term results of right-sided hemicolectomy that were performed due to malignant tumours with the comparison of the two methods. Results: A cohort of 184 right-sided hemicolectomy were performed from 01.01.2018 to 31.12.2019 from which 122 were operated on because of a malignant disease. 51 open and 71 laparoscopic operations were performed. The average age of 37 men and 34 women were 70.59 and 72.14 years, respectively. 50 patients underwent extracorporeal (EA) anastomosis and 21 intracorporeal (IA) anastomosis. Within 30 days the number of complications were 18 in the EA group and 3 in the IA group (p = 0.067). 3 from the EA group and 1 from IA group died within 30 days (p = 0.66). The average length of stay were 9.48 days in the EA group and 6.52 days in the IA group together with the complicated cases (p = 0.001) while 6.35 days and 5.55 days without the complicated cases (p = 0.09). The average duration of operation was 147 minutes in the EA and 146.47 minutes in the IA group (p = 0.11). Conclusion: We found concordance with the literature that there are fewer complications in case of IA which might be related to shorter length of stay. There is no significant difference between the surgical times. Bearing these facts in mind, IA might be suggested for right- sided laparoscopic hemicolectomy.


2020 ◽  
Vol 13 (4) ◽  
pp. 184-190
Author(s):  
Muhammad Irfan ◽  
Abdul Rasheed Qureshi ◽  
Zeeshan Ashraf ◽  
Muhammad Amjad Ramzan ◽  
Tehmina Naeem ◽  
...  

ABSTRACT Background: Conventionally Pleural effusions are suspected by history of pleuritis, evaluated by physical signs and multiple view radiography. Trans-thoracic pleural aspiration is done and aspirated pleural fluid is considered the gold-standard for pleural effusion. Chest sonography has the advantage of having high diagnostic efficacy over radiography for the detection of pleural effusion. Furthermore, ultrasonography is free from radiation hazards, inexpensive, readily available  and feasible for use in ICU, pregnant and pediatric patients. This study aims to explore the diagnostic accuracy of trans-thoracic ultrasonography for pleural fluid detection, which is free of such disadvantages. The objective is to determine the diagnostic efficacy of trans-thoracic ultrasound for detecting pleural effusion and also to assess its suitability for being a non-invasive gold-standard.   Subject and Methods: This retrospective study of 4597 cases was conducted at pulmonology  OPD-Gulab Devi Teaching Hospital, Lahore from November 2016 to July 2018. Adult patients with clinical features suggesting pleural effusions were included while those where no suspicion of pleural effusion, patients < 14 years and pregnant ladies were excluded. Patients were subjected to chest x-ray PA and Lateral views and chest ultrasonography was done by a senior qualified radiologist in OPD. Ultrasound-guided pleural aspiration was done in OPD & fluid was sent for analysis. At least 10ml aspirated fluid was considered as diagnostic for pleural effusion. Patient files containing history, physical examination, x-ray reports, ultrasound reports, pleural aspiration notes and informed consent were retrieved, reviewed and findings were recorded in the preformed proforma. Results were tabulated and conclusion was drawn by statistical analysis. Results: Out of 4597 cases, 4498 pleural effusion were manifested on CXR and only 2547(56.62%) pleural effusions were proved by ultrasound while 2050 (45.57%) cases were reported as no Pleural effusion. Chest sonography demonstrated sensitivity, specificity, PPV, NPV and diagnostic accuracy 100 % each. Conclusions: Trans-thoracic ultrasonography revealed an excellent efficacy that is why it can be considered as non-invasive gold standard for the detection of pleural effusion.


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