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1.
Front Cell Infect Microbiol ; 14: 1236630, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38435306

RESUMEN

Fusobacterium necrophorum (F. necrophorum) infection is rare in pediatrics. In addition, the detection time of F. necrophorum by blood culture is long, and the positive rate is low. Infection with F. necrophorum bacilli usually follows rapid disease progression, resulting in high mortality. In previous reports of F. necrophorum-related cases, the most dangerous moment of the disease occurred after the appearance of Lemierre's syndrome. We report an atypical case of a 6-year-old female patient who developed septic shock within 24 h of admission due to F. necrophorum infection in the absence of Lemierre's syndrome. F. necrophorum was identified in a blood sample by metagenomics next-generation sequencing (mNGS) but not by standard blood culture. The patient was finally cured and discharged after receiving timely and effective targeted anti-infection treatment. In the present case study, it was observed that the heightened virulence and invasiveness of F. necrophorum contribute significantly to its role as a primary pathogen in pediatric septic shock. This can precipitate hemodynamic instability and multiple organ failure, even in the absence of Lemierre's syndrome. The use of mNGS can deeply and rapidly identify infectious pathogens, guide the use of targeted antibiotics, and greatly improve the survival rate of patients.


Asunto(s)
Síndrome de Lemierre , Choque Séptico , Femenino , Humanos , Niño , Choque Séptico/diagnóstico , Fusobacterium necrophorum/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Antibacterianos/uso terapéutico
2.
Math Biosci Eng ; 21(2): 2608-2625, 2024 Jan 18.
Artículo en Inglés | MEDLINE | ID: mdl-38454698

RESUMEN

In the drug discovery process, time and costs are the most typical problems resulting from the experimental screening of drug-target interactions (DTIs). To address these limitations, many computational methods have been developed to achieve more accurate predictions. However, identifying DTIs mostly rely on separate learning tasks with drug and target features that neglect interaction representation between drugs and target. In addition, the lack of these relationships may lead to a greatly impaired performance on the prediction of DTIs. Aiming at capturing comprehensive drug-target representations and simplifying the network structure, we propose an integrative approach with a convolution broad learning system for the DTI prediction (ConvBLS-DTI) to reduce the impact of the data sparsity and incompleteness. First, given the lack of known interactions for the drug and target, the weighted K-nearest known neighbors (WKNKN) method was used as a preprocessing strategy for unknown drug-target pairs. Second, a neighborhood regularized logistic matrix factorization (NRLMF) was applied to extract features of updated drug-target interaction information, which focused more on the known interaction pair parties. Then, a broad learning network incorporating a convolutional neural network was established to predict DTIs, which can make classification more effective using a different perspective. Finally, based on the four benchmark datasets in three scenarios, the ConvBLS-DTI's overall performance out-performed some mainstream methods. The test results demonstrate that our model achieves improved prediction effect on the area under the receiver operating characteristic curve and the precision-recall curve.


Asunto(s)
Descubrimiento de Drogas , Redes Neurales de la Computación , Descubrimiento de Drogas/métodos , Curva ROC
3.
Infect Drug Resist ; 16: 7239-7245, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38023405

RESUMEN

Background: Rickettsia felis is a kind of zoonotic pathogen. Rickettsia felis infections of the central nervous system are rare with only a few cases reported worldwide. The early diagnosis of R. felis is difficult due to its nonspecific clinical features and laboratory tests. Here, we report two meningitis cases caused by R. felis using metagenomic next-generation sequencing (mNGS). Methods: The clinical data of patients with meningitis who were diagnosed to have R. felis through cerebrospinal fluid culture, nuclear magnetic imaging, mNGS detection from January 2019 to December 2019 in The First Clinical Hospital of Shanxi Medical University, were retrospectively analyzed, and their clinical characteristics and disease regression findings were summarized. Case Presentation: The first case was a female patient aged 23 years who was admitted to our hospital presenting with symptoms of headache, fever, and weakness in both lower limbs. Upon examination of spinal imaging, myelitis was diagnosed. However, routine examination and culture of cerebrospinal fluid did not identify the pathogen responsible. Subsequently, metagenomic second-generation sequencing (mNGS) revealed that the infection was caused by R. felis. The patient responded well to standard treatment and showed signs of recovery. The second case was a male patient aged 29 years who was admitted to our hospital with a headache and fever that had persisted for 4 days within a month. Routine examination and culture of the cerebrospinal fluid did not reveal any identifiable pathogens. However, metagenomic second-generation sequencing (mNGS) determined that the patient had a Rickettsial infection likely transmitted by a cat. The patient showed significant improvement after 14 days of doxycycline treatment. Tests for herpes simplex virus, cytomegalovirus, Epstein-Barr virus and tubercle bacillus nucleic acid in the CSF and blood were negative.Therefore mNGS of the cerebrospinal fluid was used, which identified the pathogen as R. felis. One case was diagnosed as subacute meningitis with immune-associated myelitis and the other as subacute meningitis. Conclusion: mNGS of cerebrospinal fluid can be used as a fast and effective method to identify intracranial R. felis infections.

4.
Brief Bioinform ; 24(6)2023 09 22.
Artículo en Inglés | MEDLINE | ID: mdl-37930030

RESUMEN

Bacterial infections often involve virulence factors that play a crucial role in the pathogenicity of bacteria. Accurate detection of virulence factor genes (VFGs) is essential for precise treatment and prognostic management of hypervirulent bacterial infections. However, there is a lack of rapid and accurate methods for VFG identification from the metagenomic data of clinical samples. Here, we developed a Reads-based Virulence Factors Scanner (RVFScan), an innovative user-friendly online tool that integrates a comprehensive VFG database with similarity matrix-based criteria for VFG prediction and annotation using metagenomic data without the need for assembly. RVFScan demonstrated superior performance compared to previous assembly-based and read-based VFG predictors, achieving a sensitivity of 97%, specificity of 98% and accuracy of 98%. We also conducted a large-scale analysis of 2425 clinical metagenomic datasets to investigate the utility of RVFScan, the species-specific VFG profiles and associations between VFGs and virulence phenotypes for 24 important pathogens were analyzed. By combining genomic comparisons and network analysis, we identified 53 VFGs with significantly higher abundances in hypervirulent Klebsiella pneumoniae (hvKp) than in classical K. pneumoniae. Furthermore, a cohort of 1256 samples suspected of K. pneumoniae infection demonstrated that RVFScan could identify hvKp with a sensitivity of 90%, specificity of 100% and accuracy of 98.73%, with 90% of hvKp samples consistent with clinical diagnosis (Cohen's kappa, 0.94). RVFScan has the potential to detect VFGs in low-biomass and high-complexity clinical samples using metagenomic reads without assembly. This capability facilitates the rapid identification and targeted treatment of hvKp infections and holds promise for application to other hypervirulent pathogens.


Asunto(s)
Infecciones Bacterianas , Factores de Virulencia , Humanos , Factores de Virulencia/genética , Metagenoma , Virulencia/genética , Klebsiella pneumoniae/genética , Infecciones Bacterianas/genética
5.
Front Cell Infect Microbiol ; 13: 1269963, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37965251

RESUMEN

Background: Cryptosporidium infections in humans typically result in symptoms such as abdominal pain and diarrhea. When the diarrhea is severe, it can cause serious complications and even be life-threatening, especially in patients with compromised immune systems. Case presentation: Here, we reported the use of metagenomic next-generation sequencing (mNGS) to assist in the diagnosis and treatment of a 10-year-old boy with severe Cryptosporidium infection. Despite the absence of any history of immunocompromise, the infection still resulted in severe symptoms, including shock, as well as damage to his pancreas and kidneys. The mNGS tests detected the presence of Cryptosporidium parvum when conventional methods failed. The patient received anti-parasite treatment along with supportive care to manage the condition. With disease surveillance based on regular clinical tests and sequential mNGS tests, the child recovered from the severe conditions. Conclusion: Our study emphasized the importance of recognizing the potential severity of Cryptosporidium infection, even among individuals with normal immune systems. Timely diagnosis and ongoing monitoring are essential for patient prognosis.


Asunto(s)
Criptosporidiosis , Cryptosporidium , Masculino , Niño , Humanos , Criptosporidiosis/epidemiología , Cryptosporidium/genética , Diarrea/diagnóstico , Diarrea/epidemiología , Secuenciación de Nucleótidos de Alto Rendimiento , Unidades de Cuidado Intensivo Pediátrico
6.
BMC Plant Biol ; 23(1): 415, 2023 Sep 08.
Artículo en Inglés | MEDLINE | ID: mdl-37684590

RESUMEN

As one of the largest plant specific transcription factor families, NAC family members play an important role in plant growth, development and stress resistance. To investigate the function of NAC transcription factors during abiotic stress, as well as during somatic embryogenesis, we identified and characterized the NAC gene family in Liriodendron chinense. We found that most LcNAC members contain more than three exons, with a relatively conserved gene and motif structure, especially at the N-terminus. Interspecies collinearity analysis revealed a closer relationship between the L. chinense NACs and the P. trichocarpa NACs. We analyzed the expression of LcNAC in different tissues and under three abiotic stresses. We found that 12 genes were highly expressed during the ES3 and ES4 stages of somatic embryos, suggesting that they are involved in the development of somatic embryos. 6 LcNAC genes are highly expressed in flower organs. The expression pattern analysis of LcNACs based on transcriptome data and RT-qPCR obtained from L. chinense leaves indicated differential expression responses to drought, cold, and heat stress. Genes in the NAM subfamily expressed differently during abiotic stress, and LcNAC6/18/41/65 might be the key genes in response to abiotic stress. LcNAC6/18/41/65 were cloned and transiently transformed into Liriodendron protoplasts, where LcNAC18/65 was localized in cytoplasm and nucleus, and LcNAC6/41 was localized only in nucleus. Overall, our findings suggest a role of the NAC gene family during environmental stresses in L. chinense. This research provides a basis for further study of NAC genes in Liriodendron chinense.


Asunto(s)
Liriodendron , Acetilcisteína , Núcleo Celular , Citoplasma
7.
Front Pediatr ; 11: 1183601, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37435170

RESUMEN

Background: The diagnosis of Pneumocystis pneumonia (PCP) remains challenging in certain specific clinical situations. Metagenomic next-generation sequencing (mNGS), as a novel diagnostic method, may help in the diagnosis of PCP. Case presentation: A 6-month-old male child developed acute pneumonia and sepsis. This child had previously suffered from Escherichia coli septicemia and was cured. However, the fever and dyspnea relapsed. Blood tests revealed a low lymphocyte count (0.69 × 109/L) and acute inflammatory markers such as high-level procalcitonin (8.0 ng/ml) and C-reactive protein (19 mg/dl). Chest imaging showed inflammation and decreased translucency in both lungs but no thymus shadow. Various serology tests, the 1,3-beta-D-glucan test, culture, as well as sputum smear failed to detect any pathogens. mNGS with blood helped identify 133 specific nucleic acid sequences of Pneumocystis jirovecii, suggesting an infection with this pathogen. After treatment with trimethoprim-sulfamethoxazole for 5 days, the patient's condition improved, but the child still needed ventilator support. Unfortunately, the child died soon after because of respiratory failure after his parents decided to abandon treatment. The family declined an autopsy on the child, and therefore, an anatomical diagnosis could not be obtained. Whole-exome sequencing suggested X-linked immunodeficiency. A hemizygous mutation of c.865c > t (p.r289*) was detected in the IL2RG gene, which was inherited from the mother (heterozygous state). Conclusion: This case report highlights the value of mNGS in diagnosing PCP when conventional diagnostic methods fail to identify the agent. Early onset of recurrent infectious diseases may indicate the presence of an immunodeficiency disease, for which timely genetic analysis and diagnosis are crucial.

8.
Plants (Basel) ; 12(12)2023 Jun 16.
Artículo en Inglés | MEDLINE | ID: mdl-37375974

RESUMEN

The Liriodendron chinense in the Magnoliaceae family is an endangered tree species useful for its socio-economic and ecological benefits. Abiotic stresses (cold, heat, and drought stress), among other factors, affect its growth, development, and distribution. However, GATA transcription factors (TFs) respond to various abiotic stresses and play a significant role in plant acclimatization to abiotic stresses. To determine the function of GATA TFs in L. chinense, we investigated the GATA genes in the genome of L. chinense. In this study, a total of 18 GATA genes were identified, which were randomly distributed on 12 of the total 17 chromosomes. These GATA genes clustered together in four separate groups based on their phylogenetic relationships, gene structures, and domain conservation arrangements. Detailed interspecies phylogenetic analyses of the GATA gene family demonstrated a conservation of the GATAs and a probable diversification that prompted gene diversification in plant species. In addition, the LcGATA gene family was shown to be evolutionarily closer to that of O. sativa, giving an insight into the possible LcGATA gene functions. Investigations of LcGATA gene duplication showed four gene duplicate pairs by the segmental duplication event, and these genes were a result of strong purified selection. Analysis of the cis-regulatory elements demonstrated a significant representation of the abiotic stress elements in the promoter regions of the LcGATA genes. Additional gene expressions through transcriptome and qPCR analyses revealed a significant upregulation of LcGATA17, and LcGATA18 in various stresses, including heat, cold, and drought stress in all time points analyzed. We concluded that the LcGATA genes play a pivotal role in regulating abiotic stress in L. chinense. In summary, our results provide new insights into understanding of the LcGATA gene family and their regulatory functions during abiotic stresses.

9.
Micromachines (Basel) ; 13(11)2022 Nov 19.
Artículo en Inglés | MEDLINE | ID: mdl-36422459

RESUMEN

The soft gripper has received extensive attention, due to its good adaptability and flexibility. The dielectric elastomer (DE) actuator as a flexible electroactive polymer that provides a new approach for soft grippers. However, they have the disadvantage of having a poor rigidity. Therefore, the optimization design method of a rigid-flexible soft finger is presented to improve the rigidity of the soft finger. We analyzed the interaction of the rigid and soft materials, using the finite element method (FEM), and researched the influence of the parameters (compression of the spring and pre-stretching ratio of the DE) on the bending angle. The optimal parameters were obtained using the FEM. We experimentally verified the accuracy of the proposed method. The maximum bending angle is 19.66°. Compared with the theoretical result, the maximum error is 3.84%. Simultaneously, the soft gripper with three fingers can grasp various objects and the maximum grasping quality is 11.21 g.

10.
Rev Assoc Med Bras (1992) ; 68(12): 1663-1667, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36449790

RESUMEN

OBJECTIVE: The aim of this study was to investigate the value of next-generation sequencing for the diagnosis of Streptococcus suis meningitis. METHODS: Patients with meningitis in the Department of Neurology of the Hainan General Hospital were recruited and divided into a next-generation sequencing group and a control group. In the next-generation sequencing group, we used the next-generation sequencing method to detect the specific pathogenic bacteria in the patients. In the control group, we used the cerebrospinal fluid bacterial culture method to detect the specific pathogenic bacteria in the patients. RESULTS: A total of 28 participants were recruited for this study, with 14 participants in each group. The results showed similarities in both the average age and average course of the disease between the two groups (p>0.05). The white blood cell count, percentage of neutrophils, and level of C-reactive protein in the next-generation sequencing group were significantly higher than those in the control group (p<0.05). There were similarities in both the temperature and intracranial pressure between the two groups (p>0.05). In the next-generation sequencing group, all patients (100%) were detected as having had the S. suis meningitis infection by next-generation sequencing, while only 6 (43%) patients in the control group had been detected as having the S. suis meningitis infection by cerebrospinal fluid bacterial culture. CONCLUSIONS: The positive detection rate of S. suis by the next-generation sequencing method was significantly higher compared with using a cerebrospinal fluid bacterial culture. Therefore, the next-generation sequencing method is valuable for the diagnosis of S. suis meningitis and is worthy of clinical application.


Asunto(s)
Meningitis Bacterianas , Infecciones Estreptocócicas , Streptococcus suis , Humanos , Streptococcus suis/genética , Meningitis Bacterianas/diagnóstico , Meningitis Bacterianas/microbiología , Infecciones Estreptocócicas/diagnóstico , Neutrófilos , Secuenciación de Nucleótidos de Alto Rendimiento
11.
Molecules ; 27(16)2022 Aug 12.
Artículo en Inglés | MEDLINE | ID: mdl-36014371

RESUMEN

Nowadays, drug-target interactions (DTIs) prediction is a fundamental part of drug repositioning. However, on the one hand, drug-target interactions prediction models usually consider drugs or targets information, which ignore prior knowledge between drugs and targets. On the other hand, models incorporating priori knowledge cannot make interactions prediction for under-studied drugs and targets. Hence, this article proposes a novel dual-network integrated logistic matrix factorization DTIs prediction scheme (Ro-DNILMF) via a knowledge graph embedding approach. This model adds prior knowledge as input data into the prediction model and inherits the advantages of the DNILMF model, which can predict under-studied drug-target interactions. Firstly, a knowledge graph embedding model based on relational rotation (RotatE) is trained to construct the interaction adjacency matrix and integrate prior knowledge. Secondly, a dual-network integrated logistic matrix factorization prediction model (DNILMF) is used to predict new drugs and targets. Finally, several experiments conducted on the public datasets are used to demonstrate that the proposed method outperforms the single base-line model and some mainstream methods on efficiency.


Asunto(s)
Reposicionamiento de Medicamentos , Reconocimiento de Normas Patrones Automatizadas , Algoritmos , Sistemas de Liberación de Medicamentos , Interacciones Farmacológicas , Modelos Logísticos
12.
Sci Rep ; 12(1): 7394, 2022 May 05.
Artículo en Inglés | MEDLINE | ID: mdl-35513554

RESUMEN

Intelligent recognition of assembly behaviors of workshop production personnel is crucial to improve production assembly efficiency and ensure production safety. This paper proposes a graph convolutional network model for assembly behavior recognition based on attention mechanism and multi-scale feature fusion. The proposed model learns the potential relationship between assembly actions and assembly tools for recognizing assembly behaviors. Meanwhile, the introduction of an attention mechanism helps the network to focus on the key information in assembly behavior images. Besides, the multi-scale feature fusion module is introduced to enable the network to better extract image features at different scales. This paper constructs a data set containing 15 types of workshop production behaviors, and the proposed assembly behavior recognition model is tested on this data set. The experimental results show that the proposed model achieves good recognition results, with an average assembly recognition accuracy of 93.1%.

13.
PLoS Genet ; 18(4): e1010137, 2022 04.
Artículo en Inglés | MEDLINE | ID: mdl-35421082

RESUMEN

Viral infections can alter host transcriptomes by manipulating host splicing machinery. Despite intensive transcriptomic studies on SARS-CoV-2, a systematic analysis of alternative splicing (AS) in severe COVID-19 patients remains largely elusive. Here we integrated proteomic and transcriptomic sequencing data to study AS changes in COVID-19 patients. We discovered that RNA splicing is among the major down-regulated proteomic signatures in COVID-19 patients. The transcriptome analysis showed that SARS-CoV-2 infection induces widespread dysregulation of transcript usage and expression, affecting blood coagulation, neutrophil activation, and cytokine production. Notably, CD74 and LRRFIP1 had increased skipping of an exon in COVID-19 patients that disrupts a functional domain, which correlated with reduced antiviral immunity. Furthermore, the dysregulation of transcripts was strongly correlated with clinical severity of COVID-19, and splice-variants may contribute to unexpected therapeutic activity. In summary, our data highlight that a better understanding of the AS landscape may aid in COVID-19 diagnosis and therapy.


Asunto(s)
COVID-19 , Empalme Alternativo/genética , COVID-19/genética , Prueba de COVID-19 , Humanos , Proteómica , SARS-CoV-2/genética , Transcriptoma
14.
Int J Mol Sci ; 23(3)2022 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-35163471

RESUMEN

Cold stress limits plant geographical distribution and influences plant growth, development, and yields. Plants as sessile organisms have evolved complex biochemical and physiological mechanisms to adapt to cold stress. These mechanisms are regulated by a series of transcription factors and proteins for efficient cold stress acclimation. It has been established that the ICE-CBF-COR signaling pathway in plants regulates how plants acclimatize to cold stress. Cold stress is perceived by receptor proteins, triggering signal transduction, and Inducer of CBF Expression (ICE) genes are activated and regulated, consequently upregulating the transcription and expression of the C-repeat Binding Factor (CBF) genes. The CBF protein binds to the C-repeat/Dehydration Responsive Element (CRT/DRE), a homeopathic element of the Cold Regulated genes (COR gene) promoter, activating their transcription. Transcriptional regulations and post-translational modifications regulate and modify these entities at different response levels by altering their expression or activities in the signaling cascade. These activities then lead to efficient cold stress tolerance. This paper contains a concise summary of the ICE-CBF-COR pathway elucidating on the cross interconnections with other repressors, inhibitors, and activators to induce cold stress acclimation in plants.


Asunto(s)
Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Respuesta al Choque por Frío , Regulación de la Expresión Génica de las Plantas , Fenómenos Fisiológicos de las Plantas , Procesamiento Proteico-Postraduccional , Transducción de Señal , Transactivadores , Activación Transcripcional
15.
Rev. Assoc. Med. Bras. (1992, Impr.) ; 68(12): 1663-1667, 2022. graf
Artículo en Inglés | LILACS-Express | LILACS | ID: biblio-1422551

RESUMEN

SUMMARY OBJECTIVE: The aim of this study was to investigate the value of next-generation sequencing for the diagnosis of Streptococcus suis meningitis. METHODS: Patients with meningitis in the Department of Neurology of the Hainan General Hospital were recruited and divided into a next-generation sequencing group and a control group. In the next-generation sequencing group, we used the next-generation sequencing method to detect the specific pathogenic bacteria in the patients. In the control group, we used the cerebrospinal fluid bacterial culture method to detect the specific pathogenic bacteria in the patients. RESULTS: A total of 28 participants were recruited for this study, with 14 participants in each group. The results showed similarities in both the average age and average course of the disease between the two groups (p>0.05). The white blood cell count, percentage of neutrophils, and level of C-reactive protein in the next-generation sequencing group were significantly higher than those in the control group (p<0.05). There were similarities in both the temperature and intracranial pressure between the two groups (p>0.05). In the next-generation sequencing group, all patients (100%) were detected as having had the S. suis meningitis infection by next-generation sequencing, while only 6 (43%) patients in the control group had been detected as having the S. suis meningitis infection by cerebrospinal fluid bacterial culture. CONCLUSIONS: The positive detection rate of S. suis by the next-generation sequencing method was significantly higher compared with using a cerebrospinal fluid bacterial culture. Therefore, the next-generation sequencing method is valuable for the diagnosis of S. suis meningitis and is worthy of clinical application.

16.
Microbiol Spectr ; 9(3): e0076921, 2021 12 22.
Artículo en Inglés | MEDLINE | ID: mdl-34908504

RESUMEN

Cerebrospinal fluid (CSF) circulating in the human central nervous system has long been considered aseptic in healthy individuals, because normally, the blood-brain barrier can protect against microbial invasions. However, this dogma has been called into question by several reports that microbes were identified in human brains, raising the question of whether there is a microbial community in the CSF of healthy individuals without neurological diseases. Here, we collected CSF samples and other samples, including one-to-one matched oral and skin swab samples (positive controls), from 23 pregnant women aged between 23 and 40 years. Normal saline samples (negative controls), sterile swabs, and extraction buffer samples (contamination controls) were also collected. Twelve of the CSF specimens were also used to evaluate the physiological activities of detected microbes. Metagenomic and metatranscriptomic sequencing was performed in these 116 specimens. A total of 620 nonredundant microbes were detected, which were dominated by bacteria (74.6%) and viruses (24.2%), while in CSF samples, metagenomic sequencing found only 26 nonredundant microbes, including one eukaryote, four bacteria, and 21 viruses (mostly bacteriophages). The beta diversity of microbes compared between CSF metagenomic samples and other types of samples (except negative controls) was significantly different from that of the CSF self-comparison. In addition, there was no active or viable microbe in the matched metagenomic and metatranscriptomic sequencing of CSF specimens after subtracting those also found in normal saline, DNA extraction buffer, and skin swab specimens. In conclusion, our results showed no strong evidence of a colonized microbial community present in the CSF of healthy individuals. IMPORTANCE The microbiome is prevalent throughout human bodies, with profound health implications. However, it remains unclear whether it is present and active in human CSF, which has been long considered aseptic due to the blood-brain barrier. Here, we applied unbiased metagenomic and metatranscriptomic sequencing to detect the presence of a microbiome in CSF collected from 23 pregnant women with matched controls. Analysis of 116 specimens found no strong evidence to support the presence of a colonized microbiome in CSF. Our findings will strengthen our understanding of the internal environment of the CSF in healthy people, which has strong implications for human health, especially for neurological infections and disorders, and will help further disease diagnostics, prevention, and therapeutics in clinical settings.


Asunto(s)
Bacterias/aislamiento & purificación , Bacteriófagos/aislamiento & purificación , Líquido Cefalorraquídeo/microbiología , Adulto , Bacterias/clasificación , Bacterias/genética , Barrera Hematoencefálica/microbiología , ADN Bacteriano/líquido cefalorraquídeo , ADN Viral/líquido cefalorraquídeo , Femenino , Voluntarios Sanos , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Metagenoma/genética , Metagenómica , Microbiota , Embarazo
17.
EBioMedicine ; 74: 103649, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34814051

RESUMEN

BACKGROUND: Shotgun metagenomics has been used clinically for diagnosing infectious diseases. However, most technical assessments have been limited to individual sets of reference standards, experimental workflows, and laboratories. METHODS: A reference panel and performance metrics were designed and used to examine the performance of shotgun metagenomics at 17 laboratories in a coordinated collaborative study. We comprehensively assessed the reliability, key performance determinants, reproducibility, and quantitative potential. FINDINGS: Assay performance varied significantly across sites and microbial classes, with a read depth of 20 millions as a generally cost-efficient assay setting. Results of mapped reads by shotgun metagenomics could indicate relative and intra-site (but not absolute or inter-site) microbial abundance. INTERPRETATION: Assay performance was significantly impacted by the microbial type, the host context, and read depth, which emphasizes the importance of these factors when designing reference reagents and benchmarking studies. Across sites, workflows and platforms, false positive reporting and considerable site/library effects were common challenges to the assay's accuracy and quantifiability. Our study also suggested that laboratory-developed shotgun metagenomics tests for pathogen detection should aim to detect microbes at 500 CFU/mL (or copies/mL) in a clinically relevant host context (10^5 human cells/mL) within a 24h turn-around time, and with an efficient read depth of 20M. FUNDING: This work was supported by National Science and Technology Major Project of China (2018ZX10102001).


Asunto(s)
Bacterias/aislamiento & purificación , Enfermedades Transmisibles/diagnóstico , Hongos/aislamiento & purificación , Metagenómica/instrumentación , Metagenómica/métodos , Bacterias/clasificación , Bacterias/genética , Benchmarking , China , Hongos/clasificación , Hongos/genética , Células HeLa , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Laboratorios , Metagenómica/normas , Reproducibilidad de los Resultados , Análisis de Secuencia de ADN , Flujo de Trabajo
18.
Front Cell Infect Microbiol ; 11: 541092, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33777827

RESUMEN

Background: Metagenomic next-generation sequencing (mNGS) is a powerful method for pathogen detection. In this study, we assessed the value of mNGS for bronchoalveolar lavage (BAL) samples in the diagnosis of pulmonary infections. Methods: From February 2018 to April 2019, BAL samples were collected from 235 patients with suspected pulmonary infections. mNGS and microbial culture were performed to evaluate the effectiveness of mNGS in pulmonary infection diagnosis. Results: We employed mNGS to evaluate the alpha diversity, results suggesting that patients with confirmed pathogens had a lower microbial diversity index compared to that of patients with uncertain pathogens. For the patients admitted to the respiratory intensive care unit (RICU) or on a ventilator, they experienced a lower diversity index than that of the patients in the general ward or not on a ventilator. In addition, mNGS of BAL had a diagnostic sensitivity of 88.89% and a specificity of 14.86% in pulmonary infection, with 21.16% positive predictive value (PPV) and 83.87% negative predictive value (NPV). When rare pathogens were excluded, the sensitivity of mNGS decreased to 73.33%, and the specificity increased to 41.71%. For patients in the simple pulmonary infection group and the immunocompromised group, the main infection types were bacterial infection (58.33%) and mixed-infection (43.18%). Furthermore, mNGS had an advantage over culture in describing polymicrobial ecosystem, demonstrating the microbial distribution and the dominant strains of the respiratory tract in patients with different underlying diseases. Conclusions: The study indicated that mNGS of BAL samples could provide more accurate diagnostic information in pulmonary infections and demonstrate the changes of respiratory microbiome in different underlying diseases. This method might play an important role in the clinical use of antimicrobial agents in the future.


Asunto(s)
Ecosistema , Metagenómica , Lavado Broncoalveolar , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Sensibilidad y Especificidad
19.
Mitochondrial DNA B Resour ; 6(1): 66-68, 2021 Jan 11.
Artículo en Inglés | MEDLINE | ID: mdl-33521269

RESUMEN

The first complete chloroplast genome of Morella cerifera was obtained by illumina platform sequencing technology in this study. The size of genome is 158,943 base pairs, consist of a pair of IRs 26,043 bp in length, the LSC region of 88,167 bp and SSC region of 18,690 bp. The genome has 112 unique genes, among which 79 protein-coding genes, 29 tRNAs, and 4 rRNAs. Phylogenetic analysis revealed that M. cerifera clustered with M. rubra within Myricaceae.

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