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Ceftriaxone-resistant Neisseria gonorrhoeae FC428-like strains have disseminated across the Asia-Pacific region, with a continuous rise in prevalence during 2015-2022. To mitigate the effect of these strains, we advocate for enhanced molecular diagnostics, expanded surveillance networks, and a regionally coordinated effort to combat the global spread of FC428-like strains.
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Antibacterianos , Ceftriaxona , Farmacorresistencia Bacteriana , Gonorrea , Neisseria gonorrhoeae , Neisseria gonorrhoeae/efectos de los fármacos , Neisseria gonorrhoeae/genética , Ceftriaxona/farmacología , Humanos , Gonorrea/microbiología , Gonorrea/epidemiología , Gonorrea/tratamiento farmacológico , Asia/epidemiología , Antibacterianos/farmacología , Pruebas de Sensibilidad Microbiana , Prevalencia , Historia del Siglo XXIRESUMEN
BACKGROUND: Antimicrobial resistance in gonorrhea has become a growing global public health burden. Neisseria gonorrhoeae isolates with resistance to ceftriaxone, the last remaining first-line option, represent an emerging threat of untreatable gonorrhea. METHODS: A total of ten ceftriaxone-resistant N. gonorrhoeae FC428 isolates and two isolates harboring a novel mosaic penA-232.001 allele from 160 gonococcal isolates in Chengdu in 2019-2020 was described in the present study. Multilocus sequence typing (MLST) and N. gonorrhoeae sequence typing for antimicrobial resistance (NG-STAR) were performed to characterize the isolates. Whole genome sequencing and maximum-likelihood method were performed to infer how the genetic phylogenetic tree of these isolates looks like. Recombination analysis was performed using the RDP4 software. This study was registered in the Chinese Clinical Trial Registry (ChiCTR2100048771, registration date: 20210716). RESULTS: The genetic phylogeny showed that the ten FC428 isolates sporadically clustered into different phylogenetic clades, suggesting different introductions and local transmission of FC428. Two isolates showed close genetic relatedness to ceftriaxone-resistant clone A8806, which was only reported from Australia in 2013. Homologous recombination events were detected in penA between Neisseria gonorrhoeae and commensal Neisseria species (N. perflava and N. polysaccharea), providing evidence of commensal Neisseria species might serve as reservoirs of ceftriaxone resistance-mediating penA sequences in clinical gonococcal strains. CONCLUSIONS: Our results demonstrate further dissemination of FC428 in China and resurgence risks of sporadic ceftriaxone-resistant A8806 to become the next clone to spread.
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Antiinfecciosos , Gonorrea , Humanos , Neisseria gonorrhoeae/genética , Ceftriaxona/farmacología , Tipificación de Secuencias Multilocus , Filogenia , Programas InformáticosRESUMEN
The emerging cephalosporin-resistant Neisseria gonorrhoeae poses an urgent threat to the continued efficacy of the last-line monotherapy for gonorrhea. Consequently, high-throughput, accurate, and reasonable molecular assays are urgently needed for strengthening antimicrobial-resistance surveillance in N. gonorrhoeae. In this study, we designed a high-throughput multiplex method that incorporates high-resolution melting technology and is based on a 6-codon assay (among the most parsimonious assays) developed following comprehensive and systematic reviews. The results showed that our method can precisely distinguish specific single-nucleotide polymorphisms in resistance-associated genes with a specificity and sensitivity of 100% and a detection limit as low as 10 copies per reaction. This method can be directly applied to clinical samples without cumbersome culture and successfully predicted all cephalosporin-resistant isolates (sensitivity: 100%). The method presented here represents a technique for rapid testing of antimicrobial resistance and will serve as a valuable tool for tailor-made antimicrobial therapy and for monitoring the transmission of cephalosporin-resistant strains.
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Gonorrea , Neisseria gonorrhoeae , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Cefalosporinas/farmacología , Cefalosporinas/uso terapéutico , Codón , Gonorrea/diagnóstico , Humanos , Pruebas de Sensibilidad MicrobianaRESUMEN
The growing antimicrobial resistance (AMR) in Neisseria gonorrhoeae is a serious global threat to gonococcal therapy. Molecular typing is an ideal tool to reveal the association between specific genotype and resistance phenotype that provides effective data for tracking the transmission of resistant clones of N. gonorrhoeae In our study, we aimed to describe the molecular epidemiology of AMR and the distribution of resistance-associated genotypes in Shenzhen during 2014-2018. In total, 909 isolates were collected from Shenzhen from 2014-2018. Two typing schemes, multilocus sequence typing (MLST) and N. gonorrhoeae Sequence Typing for Antimicrobial Resistance (NG-STAR), were performed for all isolates. The distribution of resistance-associated genotypes was described using goeBURST analysis combined with data of logistic regression. Among 909 isolates, ST8123, ST7363, ST1901, ST7365, and ST7360 were most the common MLST sequence types (STs), and ST348, ST2473, ST497, and ST199 were the most prevalent NG-STAR STs. The logistic regression analysis showed that NG-STARST497, MLSTST7365, and MLSTST7360 were typically associated with decreased susceptibility to ceftriaxone. Furthermore, the internationally spreading ESC-resistant clone MLSTST1901 has been prevalent at least in 2014 in Shenzhen and showed a significant increase during 2014-2018. Additionally, MLSTST7363 owns the potential to become the next internationally spreading ceftriaxone-resistant ST. In conclusions, we performed a comprehensive epidemiological study to explore the correlation between AMR and specific STs, which provided important data for future studies of the molecular epidemiology of AMR in N. gonorrhoeae Besides, these findings provide insight for adjusting surveillance strategies and therapy management in Shenzhen.
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BACKGROUND: Antimicrobial resistance (AMR) of Neisseria gonorrhoeae has spread worldwide. Rapid and comprehensive methods are needed to describe N. gonorrhoeae AMR profiles accurately. A method based on multiplex amplicon sequencing was developed to simultaneously sequence 13 genes related to AMR in N. gonorrhoeae directly from clinical samples. METHODS: Nine N. gonorrhoeae strains were used for the establishment and validation of the method. Eleven urethral swabs and their corresponding cultured isolates were matched as pairs to determine the accuracy of the method. Mock samples with different dilutions were prepared to determine the sensitivity of the method. Five nongonococcal Neisseria strains and 24 N. gonorrhoeae negative clinical samples were used to evaluate the cross-reactivity. Finally, the method was applied to 64 clinical samples to assess its performance. RESULTS: Using Sanger sequencing as a reference method, sequences recovered from amplicon sequencing had a base accuracy of over 99.5% and the AMR sites were correctly identified. The limit of detection (LOD) was lower than 31 copies/reaction. No significant cross-reactivity was observed. Furthermore, target genes were successfully recovered from 64 clinical samples including 9 urines, demonstrating this method could be used in different types of samples. For clinical samples, the results can be obtained within a time frame of 7 h 40 min to 10 h 40 min, while for isolates, the turnaround time was approximately 2 h shorter. CONCLUSIONS: This method can serve as a versatile and convenient culture-free diagnostic method with the advantages of high sensitivity and accuracy.
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Gonorrea , Secuenciación de Nanoporos , Antibacterianos/farmacología , Farmacorresistencia Bacteriana/genética , Gonorrea/diagnóstico , Humanos , Pruebas de Sensibilidad Microbiana , Reacción en Cadena de la Polimerasa Multiplex , Neisseria gonorrhoeae/efectos de los fármacos , Neisseria gonorrhoeae/genéticaRESUMEN
OBJECTIVES: The continuous emergence of ceftriaxone-resistant Neisseria gonorrhoeae strains threatens the effectiveness of current treatment regimens for gonorrhoea. The objective of the present study was to characterize three ceftriaxone-resistant N. gonorrhoeae strains with a novel mosaic penA allele isolated in China. METHODS: Three ceftriaxone-resistant Neisseria gonorrhoeae strains (GC150, GC161 and GC208) isolated in 2017 were characterized by N. gonorrhoeae multiantigen sequence typing (NG-MAST), MLST and N. gonorrhoeae sequence typing for antimicrobial resistance (NG-STAR). Recombination analyses were performed using the SimPlot software. RESULTS: Three strains had the same antibiotic resistance profiles, with resistance to ceftriaxone (MIC 0.5 mg/L), ciprofloxacin (MIC 8.0 mg/L), penicillin (MIC 2.0 mg/L) and tetracycline (MIC 2.0-8.0 mg/L). STs were assigned as MLST7360, NG-MAST14292 and NG-STAR1611/NG-STAR1612. The penA gene of these three strains differed from previous ceftriaxone-resistant gonococcal strains and harboured a novel mosaic allele (penA-121.001). Like N. gonorrhoeae FC428, a widely disseminated ceftriaxone-resistant strain that was initially described in Japan in 2015, all strains also possessed substitutions A311V and T483S in PBP2, which are associated with resistance to ceftriaxone. Potential recombination events were detected in penA between N. gonorrhoeae strain FC428 and commensal Neisseria species. Our results provide further evidence that the commensal Neisseria species (Neisseria cinerea and Neisseria perflava) can serve as a reservoir of ceftriaxone resistance-mediating penA sequences in clinical gonococcal strains. CONCLUSIONS: The emergence of such strains may be the result of the interspecies recombination of penA genes between N. gonorrhoeae strain FC428 and commensal Neisseria species.
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Gonorrea , Neisseria gonorrhoeae , Antibacterianos/farmacología , Ceftriaxona/farmacología , China , Humanos , Japón , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Neisseria , Neisseria gonorrhoeae/genética , Programas InformáticosRESUMEN
OBJECTIVES: Increased awareness of the international spread of the ceftriaxone-resistant Neisseria gonorrhoeae FC428 clone, which threatens recommended dual therapy, is essential. The objective of the present study was to develop and evaluate a rapid, simple and cost-effective method based on high-resolution melting (HRM) analysis for direct detection of the FC428 clone from clinical isolates and specimens. METHODS: The singleplex HRM assay was designed to identify the FC428 clone by using specific primers, which flank the alteration A311V in the penA-60.001 allele. Analytical performance was initially evaluated by testing 623 isolates and a panel of non-gonococcal strains. To ensure the method can be directly applied in clinical samples, two internal control targets (opa and porA) were also designed and included in the final multiplex HRM assay. Two hundred and eighty-two clinical samples (94 urine and 188 urethral/genital swabs) were then analysed using this multiplex HRM assay. RESULTS: The FC428 clone was easily differentiated from the non-mosaic alleles and other mosaic alleles without A311 mutations by comparing the differences in melt curves. Cross-reactivity was not observed for the penA-60.001 allele when testing 15 non-gonococcal Neisseria strains. When applied to the 623 isolates, the HRM assay successfully characterized one isolate as an FC428 clone (MLST1903, NG-MAST3435, NG-STAR233). Our data show that the multiplex HRM assay with high specificity can be directly applied in clinical samples. CONCLUSIONS: This method can generate results within 90 min at a cost of less than US$0.5 per isolate or sample, making this assay an ideal tool for large epidemiological studies to enhance surveillance of the internationally transmitted ceftriaxone-resistant N. gonorrhoeae FC428 clone.
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Antibacterianos/farmacología , Ceftriaxona/farmacología , Neisseria gonorrhoeae/efectos de los fármacos , Neisseria gonorrhoeae/genética , Alelos , Salud Global , Gonorrea/diagnóstico , Gonorrea/microbiología , Gonorrea/transmisión , Humanos , Pruebas de Sensibilidad Microbiana , Sensibilidad y Especificidad , Temperatura de TransiciónRESUMEN
BACKGROUND: Complicated mechanisms and variable determinants related to drug resistance pose a major challenge to obtain comprehensive antimicrobial resistance (AMR) profiles of Neisseria gonorrhoeae. Meanwhile, cephalosporin-resistant mosaic penA alleles have been reported worldwide. Therefore, it is urgent to monitor the expansion of cephalosporin-resistant mosaic penA alleles. OBJECTIVES: To develop a comprehensive high-throughput method to efficiently screen AMR determinants. METHODS: We developed a method based on multiplex PCR with MALDI-TOF MS, which can simultaneously screen for 24 mutations associated with multiple antimicrobial agents in 19 gonococcal AMR loci (NG-AMR-MS). The performance of the NG-AMR-MS method was assessed by testing 454 N. gonorrhoeae isolates with known MICs of six antibiotics, eight non-gonococcal Neisseria strains, 214 clinical samples and three plasmids with a confirmed mosaic penA allele. RESULTS: The results show that NG-AMR-MS had a specificity of 100% with a sensitivity as low as 10 copies per reaction (except for PorB A121D/N/G, 100 copies per reaction). For clinical samples with gonococcal load >5 copies/µL, the method can accurately identify 20 AMR mutations. In addition, the method successfully detected specific cephalosporin-resistant strains with the A311V mutation in the penA allele. CONCLUSIONS: Our high-throughput method can provide comprehensive AMR profiles within a multiplex format. Furthermore, the method can be directly applied to screening for AMR among clinical samples, serving as an effective tool for overall monitoring of N. gonorrhoeae AMR and also provides a powerful means to comprehensively improve the level of monitoring.
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Farmacorresistencia Bacteriana , Gonorrea , Neisseria gonorrhoeae , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Farmacorresistencia Bacteriana/efectos de los fármacos , Gonorrea/tratamiento farmacológico , Humanos , Espectrometría de Masas , Pruebas de Sensibilidad Microbiana , Neisseria gonorrhoeae/efectos de los fármacos , Neisseria gonorrhoeae/genéticaRESUMEN
The low frequency magnetic field detection ability of magnetoresistive (MR)sensor is seriously affected by 1/f noise. At present, the method to suppress the influence of low frequency noise is mainly to modulate the measured magnetic field by mechanical resonance. In this paper, a novel modulation concept employing a magnetoelectric coupling effect is proposed. A design method of modulation structure based on an equivalent magnetic circuit model (EMCM) and a single domain model of in-plane moment was established. An EMCM was established to examine the relationship between the permeability of flux modulation film (FMF) and modulation efficiency, which was further verified through a finite element simulation model (FESM). Then, the permeability modulated by the voltage of a ferroelectric/ferromagnetic (FE/FM) multiferroic heterostructure was theoretically studied. Combining these studies, the modulation structure and the material were further optimized, and a FeSiBPC/PMN-PT sample was prepared. Experimental results show that the actual magnetic susceptibility modulation ability of FeSiBPC/PMN-PT reached 150 times, and is in good agreement with the theoretical prediction. A theoretical modulation efficiency higher than 73% driven by a voltage of 10 V in FeSiBPC/PMN-PT can be obtained. These studies show a new concept for magnetoelectric coupling application, and establish a new method for magnetic field modulation with a multiferroic heterostructure.
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To improve the sensitivity of the magnetic tunnel junction(MTJ)sensor, a novel architecture for a double-gap magnetic flux concentrator (MFC) was studied theoretically and experimentally in this paper. The three-dimensional finite element model of magnetic flux was established to optimize the magnetic field amplification factor, with different gaps. The simulation results indicate that the sensitivity of an MTJ sensor with a double-gap MFC can be significantly better than that of a sensor with a traditional single-gap MFC, due to the fact that the magnetic magnification sharply increases with the decrease in effective gap width. Besides this, the half-bridge MTJ sensors with the double-gap MFC were fabricated using photolithography, ion milling, evaporation, and electroplating processes. Experimental results show that the sensitivity of the MTJ sensor increased by ten times compared to the sensor without the double-gap MFC, which underlines the theoretical predictions. Furthermore, there is no significant increase in the sensor noise. The work in this paper contributes to the development of high-performance MTJ sensors.
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Superconducting and topological states are two most intriguing quantum phenomena in solid materials. The entanglement of these two states, the topological superconducting state, will give rise to even more exotic quantum phenomena. While many materials are found to be either a superconductor or a topological insulator, it is very rare that both states exist in one material. Here, we demonstrate by first-principles theory as well as scanning tunnelling spectroscopy and angle-resolved photoemission spectroscopy experiments that the recently discovered 'two-dimensional (2D) superconductor' of single-layer FeSe also exhibits 1D topological edge states within an energy gap of â¼40 meV at the M point below the Fermi level. It is the first 2D material that supports both superconducting and topological states, offering an exciting opportunity to study 2D topological superconductors through the proximity effect.
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BACKGROUND: Cavernous hemangiomas (CHs) of the adrenal gland are extremely rare. To date, only a few studies of adrenal CH imaging have been reported. PURPOSE: To analyze the computed tomography (CT) imaging findings of adrenal CHs. MATERIAL AND METHODS: Ten cases of adrenal CHs confirmed by a histopathological examination were retrospectively analyzed. All of the patients had undergone unenhanced and enhanced CT examinations, and eight had also undergone multiphase CT enhancement examinations. CT characteristics, including shape, size, margin, attenuation, and enhancement patterns, were analyzed. RESULTS: The study included six women and four men with a mean age of 49.2 years (age range, 25-62 years) and no signs of abnormal endocrine activity. The unenhanced CTs showed well-defined, heterogeneous (n = 8) or homogeneous (n = 2) density masses with scattered (n = 8) or spread calcifications (n = 2) in six tumors. In the contrast-enhanced CTs, seven tumors appeared to be marked with heterogeneous enhancement, whereas three cases exhibited no obvious enhancement. The evaluation of the pattern of dynamic enhancement in eight patients revealed that the tumors showed early peripheral enhancement (n = 4), early central enhancement (n = 1), and mixed enhancement (n = 1) with progressive partial filling-in, and no obvious enhancement in any phases (n = 2). CONCLUSION: Adrenal CHs should be included in the differential diagnosis when an adrenal neoplasm is incidentally found and appears as a well-defined, heterogeneous mass with calcifications and various enhancement patterns, including heterogeneous enhancement with characteristic progressive partial filling-in, as well as lack of enhancement in any phase.
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Neoplasias de las Glándulas Suprarrenales/diagnóstico por imagen , Hemangioma Cavernoso/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Adulto , Medios de Contraste , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
Respiratory infections continue to pose a significant threat to human health. It is important to accurately and rapidly detect respiratory viruses. To compensate for the limits of current respiratory virus detection methods, we developed a 24-plex analysis (common respiratory virus-mass spectrometry [CRV-MS]) that can simultaneously detect and identify 21 common respiratory viruses based on a matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry system. To evaluate the efficacy of the CRV-MS method, we used 102 samples that were confirmed positive for these common respiratory viruses. All tests using the CRV-MS method were effective, with no cross-reactivity observed with other common respiratory viruses. To confirm the usefulness of the CRV-MS method, we screened 336 nasal and throat swabs that were collected from adults or children with suspected viral acute respiratory tract infections using the CRV-MS method and consensus PCR/reverse transcription-PCR (RT-PCR) methods. Excluding four RNase P-negative samples, the CRV-MS and consensus PCR/RT-PCR methods detected respiratory viruses in 92.5% (307/332) and 89.5% (297/332) of the samples, respectively. The two methods yielded identical results for 306 (92.2%) samples, including negative results for 25 samples (7.5%) and positive results for 281 samples (84.6%). Differences between the two methods may reflect their different sensitivities. The CRV-MS method proved to be sensitive and robust, and it can be used in large-scale epidemiological studies of common respiratory virus infections.
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Reacción en Cadena de la Polimerasa Multiplex/métodos , Infecciones del Sistema Respiratorio/diagnóstico , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Virología/métodos , Virosis/diagnóstico , Virus/aislamiento & purificación , Adulto , Niño , Preescolar , Humanos , Nariz/microbiología , Faringe/virología , Infecciones del Sistema Respiratorio/virología , Sensibilidad y Especificidad , Factores de Tiempo , Virus/química , Virus/clasificación , Virus/genéticaRESUMEN
Understanding the mechanism of high transition temperature (T{c}) superconductivity in cuprates has been hindered by the apparent complexity of their multilayered crystal structure. Using a cryogenic scanning tunneling microscopy (STM), we report on layer-by-layer probing of the electronic structures of all ingredient planes (BiO, SrO, CuO{2}) of Bi{2}Sr{2}CaCu_2}O{8+δ} superconductor prepared by argon-ion bombardment and annealing technique. We show that the well-known pseudogap (PG) feature observed by STM is inherently a property of the BiO planes and thus irrelevant directly to Cooper pairing. The SrO planes exhibit an unexpected van Hove singularity near the Fermi level, while the CuO{2} planes are exclusively characterized by a smaller gap inside the PG. The small gap becomes invisible near T{c}, which we identify as the superconducting gap. The above results constitute severe constraints on any microscopic model for high T{c} superconductivity in cuprates.
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The recent observation of the superconducting state at atomic scale has motivated the pursuit of exotic condensed phases in two-dimensional (2D) systems. Here we report on a superconducting phase in two-monolayer crystalline Ga films epitaxially grown on wide-band-gap semiconductor GaN(0001). This phase exhibits a hexagonal structure and only 0.552 nm in thickness, nevertheless, brings about a superconducting transition temperature Tc as high as 5.4 K, confirmed by in situ scanning tunneling spectroscopy and ex situ electrical magnetotransport and magnetization measurements. The anisotropy of critical magnetic field and Berezinski-Kosterlitz-Thouless-like transition are observed, typical for the 2D superconductivity. Our results demonstrate a novel platform for exploring atomic-scale 2D superconductors, with great potential for understanding the interface superconductivity.
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Introduction: This study aims to determine the etiological, sociodemographic, and clinical characteristics of STIs, and the level of resistance in M. genitalium in Shenzhen, a representative first-tier city of southern China. Methods: A multicenter cross-sectional study was conducted and 7886 sexually active participants attending STI-related departments were involved from 22 hospitals. Nine STI-related organisms including N. gonorrhoeae, C. trachomatis, T. vaginalis, M. genitalium, HSV-1, HSV-2, M. hominis, U. parvum, and U. urealyticum were screened. Results: Being single or divorced was associated with increased detection of N. gonorrhoeae, C. trachomatis, M. genitalium, HSV-1, HSV-2 and M. hominis. Lower education level was associated with increased detection of C. trachomatis, HSV-2 and M. hominis. No insurance coverage was an independent risk factor for T. vaginalis, M. hominis and U. parvum positivity. Three resistance-determining regions related to macrolide and fluoroquinolone were sequenced in 154 M. genitalium positive samples, among which 90.3% harbored mutations related to macrolide or fluroquinolone resistance and 67.5% were multidrug-resistant M. genitalium. A2072G in 23S rRNA and Ser83Ile in parC were the most common mutations. M. hominis was associated with manifestations of bacterial vaginosis in female and epididymitis in male. Conclusions: Single or divorced individuals, those with lower education level and individuals without insurance are higher-risk key populations for STIs. The prevalence of antimicrobial-resistant M. genitalium in Shenzhen is high. Detection of M. hominis increased significantly with lower education level and no health insurance coverage, and it is associated with bacterial vaginosis or epididymitis, indicating that M. hominis deserves further attention.
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Antibacterianos , Mycoplasma genitalium , Enfermedades de Transmisión Sexual , Humanos , Estudios Transversales , China/epidemiología , Femenino , Masculino , Adulto , Mycoplasma genitalium/genética , Mycoplasma genitalium/aislamiento & purificación , Mycoplasma genitalium/efectos de los fármacos , Enfermedades de Transmisión Sexual/epidemiología , Enfermedades de Transmisión Sexual/microbiología , Adulto Joven , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Persona de Mediana Edad , Farmacorresistencia Bacteriana , Adolescente , Factores de Riesgo , Factores Socioeconómicos , Infecciones por Mycoplasma/epidemiología , Infecciones por Mycoplasma/microbiología , Pruebas de Sensibilidad Microbiana , PrevalenciaRESUMEN
We conducted a case-control study to explore the prevalence of 10 human polyomaviruses in fecal specimens from hospitalized children with diarrhea and asymptomatic control subjects by using multiplex PCR detected by matrix-assisted laser desorption ionization-time of flight mass spectrometry. The differences between cases and controls were not statistically significant.
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Heces/virología , Gastroenteritis/epidemiología , Gastroenteritis/virología , Infecciones por Polyomavirus/epidemiología , Infecciones por Polyomavirus/virología , Poliomavirus/aislamiento & purificación , Estudios de Casos y Controles , Niño Hospitalizado , Preescolar , Diarrea/epidemiología , Diarrea/virología , Femenino , Humanos , Lactante , Masculino , Reacción en Cadena de la Polimerasa Multiplex , Poliomavirus/clasificación , Prevalencia , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Human papillomavirus (HPV) is the principal cause of invasive cervical cancer and benign genital lesions. There are currently 30 HPV types linked to cervical cancer. HPV infection also leads to other types of cancer. We developed a 61-plex analysis of these 30 HPV types by examining two genes, E6 and L1, using MassARRAY matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (PCR-MS). Two hundred samples from homosexual males (HM) were screened by PCR-MS and MY09/MY11 primer set-mediated PCR (MY-PCR) followed by sequencing. One hundred thirty-five formalin-fixed, paraffin-embedded (FFPE) cervical cancer samples were also analyzed by PCR-MS, and results were compared to those of the commercially available GenoArray (GA) assay. One or more HPV types were identified in 64.5% (129/200) of the samples from HM. Comprising all 30 HPV types, PCR-MS detected 51.9% (67/129) of samples with multiple HPV types, whereas MY-PCR detected only one single HPV type in these samples. All PCR-MS results were confirmed by MY-PCR. In the cervical cancer samples, PCR-MS and GA detected 97% (131/135) and 90.4% (122/135) of HPV-positive samples, respectively. PCR-MS and GA results were fully concordant for 122 positive and 4 negative samples. The sequencing results for the 9 samples that tested negative by GA were completely concordant with the positive PCR-MS results. Multiple HPV types were identified in 25.2% (34/135) and 55.6% (75/135) of the cervical cancer samples by GA and PCR-MS, respectively, and results were confirmed by sequencing. The new assay allows the genotyping of >1,000 samples per day. It provides a good alternative to current methods, especially for large-scale investigations of multiple HPV infections and degraded FFPE samples.