RESUMO
Infectious vaginitis due to bacterial vaginosis (BV), vulvovaginal candidiasis (VVC), and Trichomonas vaginalis accounts for a significant proportion of all gynecologic visits in the United States. A prospective multicenter clinical study was conducted to validate the performance of two new in vitro diagnostic transcription-mediated amplification nucleic acid amplification tests (NAATs) for diagnosis of BV, VVC, and trichomoniasis. Patient- and clinician-collected vaginal-swab samples obtained from women with symptoms of vaginitis were tested with the Aptima BV and Aptima Candida/Trichomonas vaginitis (CV/TV) assays. The results were compared to Nugent (plus Amsel for intermediate Nugent) scores for BV, Candida cultures and DNA sequencing for VVC, and a composite of NAAT and culture for T. vaginalis The prevalences of infection were similar for clinician- and patient-collected samples: 49% for BV, 29% for VVC due to the Candida species group, 4% for VVC due to Candida glabrata, and 10% for T. vaginalis Sensitivity and specificity estimates for the investigational tests in clinician-collected samples were 95.0% and 89.6%, respectively, for BV; 91.7% and 94.9% for the Candida species group; 84.7% and 99.1% for C. glabrata; and 96.5% and 95.1% for T. vaginalis Sensitivities and specificities were similar in patient-collected samples. In a secondary analysis, clinicians' diagnoses, in-clinic assessments, and investigational-assay results were compared to gold standard reference methods. Overall, the investigational assays had higher sensitivity and specificity than clinicians' diagnoses and in-clinic assessments, indicating that the investigational assays were more predictive of infection than traditional diagnostic methods. These results provide clinical-efficacy evidence for two in vitro diagnostic NAATs that can detect the main causes of vaginitis.
Assuntos
Candidíase Vulvovaginal/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/normas , Kit de Reagentes para Diagnóstico/normas , Vaginite por Trichomonas/diagnóstico , Vaginose Bacteriana/diagnóstico , Adolescente , Adulto , Idoso , Bactérias/genética , Candida/genética , Candidíase Vulvovaginal/microbiologia , Estudos Transversais , Feminino , Humanos , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico/métodos , Estudos Prospectivos , Sensibilidade e Especificidade , Trichomonas vaginalis/genética , Estados Unidos , United States Food and Drug Administration , Vagina/microbiologia , Vaginose Bacteriana/microbiologia , Adulto JovemRESUMO
BACKGROUND: During the past decade, breakthroughs in sequencing technology and computational biology have provided the basis for studies of the myriad ways in which microbial communities ("microbiota") in and on the human body influence human health and disease. In almost every medical specialty, there is now a growing interest in accurate and replicable profiling of the microbiota for use in diagnostic and therapeutic application. CONTENT: This review provides an overview of approaches, challenges, and considerations for diagnostic applications borrowing from other areas of molecular diagnostics, including clinical metagenomics. Methodological considerations and evolving approaches for microbiota profiling from mitochondrially encoded 16S rRNA-based amplicon sequencing to metagenomics and metatranscriptomics are discussed. To improve replicability, at least the most vulnerable steps in testing workflows will need to be standardized and continuous efforts needed to define QC standards. Challenges such as purity of reagents and consumables, improvement of reference databases, and availability of diagnostic-grade data analysis solutions will require joint efforts across disciplines and with manufacturers. SUMMARY: The body of literature supporting important links between the microbiota at different anatomic sites with human health and disease is expanding rapidly and therapeutic manipulation of the intestinal microbiota is becoming routine. The next decade will likely see implementation of microbiome diagnostics in diagnostic laboratories to fully capitalize on technological and scientific advances and apply them in routine medical practice.
Assuntos
Metagenômica/métodos , Microbiota/genética , Bactérias/genética , Bactérias/isolamento & purificação , Resistência a Medicamentos/genética , Fungos/genética , Fungos/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Metagenômica/normas , Mitocôndrias/genética , Controle de Qualidade , RNA Ribossômico 16S/química , RNA Ribossômico 16S/metabolismoRESUMO
BACKGROUND: Until recently, no HPV test had been US FDA-approved for SurePath preservative. Clinical performance remains incompletely understood. The clinical performances of the Cobas HPV Test (Cobas) and Hybrid Capture 2 High-Risk HPV DNA Test (HC2) with PreservCyt and SurePath preservatives were compared. METHODS: Cervical cytology samples were collected in both preservatives in random order from women age 21+ (n = 244) referred for colposcopy. Before cytology processing and pelleting, SurePath samples were tested by the Cobas test with and without buffered SDS heat pretreatment. SurePath pellets were tested by the HC2 test and by the Cobas test (with pretreatment). Performance characteristics were calculated in relation to cases of cervical in-traepithelial neoplasia grade 2 or higher (CIN2+) as the clinical target outcome. All HPV-positive samples were also genotyped with the Linear Array test. RESULTS: CIN2+ was detected in 42 patients (17.2%). For both HPV tests, there was a trend towards higher positivity and sensitivity for SurePath compared to PreservCyt preservative. The Cobas test had higher sensitivity than HC2 and the HC2 test had higher specificity than Cobas. Pretreated SurePath samples produced results similar to untreated ones, despite a two-fold dilution during pretreatment [sensitivity %: 95.1 (82.2 - 99.2) vs. 94.3 (79.5 - 99.0); specificity %: 33.0 (26.6 - 40.1) vs. 33.0 (26.4 - 40.3)]. CONCLUSIONS: There was good agreement between the preservatives and HPV tests in detecting HPV and between the Cobas and Linear Array tests for genotyping HR-HPV. These trends were not statistically significant due to the limited number of CIN2+ cases. However, these data may help in evaluations of preservative selection for colposcopy samples. Pre-treatment for Cobas testing eliminated invalid results due to clots. The Cobas test has been FDA-approved for use with heat pretreated SurePath samples.
Assuntos
Testes de DNA para Papilomavírus Humano , Manejo de Espécimes , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Adulto JovemRESUMO
Background: Community-acquired pneumonia (CAP) is a leading cause of pediatric hospitalization. Pathogen identification fails in approximately 20% of children but is critical for optimal treatment and prevention of hospital-acquired infections. We used two broad-spectrum detection strategies to identify pathogens in test-negative children with CAP and asymptomatic controls. Methods: Nasopharyngeal/oropharyngeal (NP/OP) swabs from 70 children <5 years with CAP of unknown etiology and 90 asymptomatic controls were tested by next-generation sequencing (RNA-seq) and pan viral group (PVG) PCR for 19 viral families. Association of viruses with CAP was assessed by adjusted odds ratios (aOR) and 95% confidence intervals controlling for season and age group. Results: RNA-seq/PVG PCR detected previously missed, putative pathogens in 34% of patients. Putative viral pathogens included human parainfluenza virus 4 (aOR 9.3, P = .12), human bocavirus (aOR 9.1, P < .01), Coxsackieviruses (aOR 5.1, P = .09), rhinovirus A (aOR 3.5, P = .34), and rhinovirus C (aOR 2.9, P = .57). RNA-seq was more sensitive for RNA viruses whereas PVG PCR detected more DNA viruses. Conclusions: RNA-seq and PVG PCR identified additional viruses, some known to be pathogenic, in NP/OP specimens from one-third of children hospitalized with CAP without a previously identified etiology. Both broad-range methods could be useful tools in future epidemiologic and diagnostic studies.
Assuntos
Infecções Comunitárias Adquiridas/virologia , Metagenômica/métodos , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase/métodos , Vírus/genética , Pré-Escolar , Estudos de Coortes , Infecções Comunitárias Adquiridas/diagnóstico , Humanos , Lactente , Recém-Nascido , Pneumonia Viral/diagnóstico , Análise de Sequência de RNA/métodosRESUMO
Background: The role of human bocavirus (HBoV) in respiratory illness is uncertain. HBoV genomic DNA is frequently detected in both ill and healthy children. We hypothesized that spliced viral capsid messenger RNA (mRNA) produced during active replication might be a better marker for acute infection. Methods: As part of the Etiology of Pneumonia in the Community (EPIC) study, children aged <18 years who were hospitalized with community-acquired pneumonia (CAP) and children asymptomatic at the time of elective outpatient surgery (controls) were enrolled. Nasopharyngeal/oropharyngeal specimens were tested for HBoV mRNA and genomic DNA by quantitative polymerase chain reaction. Results: HBoV DNA was detected in 10.4% of 1295 patients with CAP and 7.5% of 721 controls (odds ratio [OR], 1.4 [95% confidence interval {CI}, 1.0-2.0]); HBoV mRNA was detected in 2.1% and 0.4%, respectively (OR, 5.1 [95% CI, 1.6-26]). When adjusted for age, enrollment month, and detection of other respiratory viruses, HBoV mRNA detection (adjusted OR, 7.6 [95% CI, 1.5-38.4]) but not DNA (adjusted OR, 1.2 [95% CI, .6-2.4]) was associated with CAP. Among children with no other pathogens detected, HBoV mRNA (OR, 9.6 [95% CI, 1.9-82]) was strongly associated with CAP. Conclusions: Detection of HBoV mRNA but not DNA was associated with CAP, supporting a pathogenic role for HBoV in CAP. HBoV mRNA could be a useful target for diagnostic testing.
Assuntos
Bocavirus/isolamento & purificação , Proteínas do Capsídeo/genética , Infecções por Parvoviridae/diagnóstico , Pneumonia Viral/diagnóstico , RNA Mensageiro/isolamento & purificação , RNA Viral/isolamento & purificação , Doença Aguda , Bocavirus/genética , Estudos de Casos e Controles , Criança , Pré-Escolar , Infecções Comunitárias Adquiridas/diagnóstico , Infecções Comunitárias Adquiridas/virologia , Hospitalização , Humanos , Lactente , Masculino , Nasofaringe/virologia , Orofaringe/virologia , Estudos Prospectivos , Manejo de EspécimesRESUMO
Ehrlichiosis is a bacterial zoonosis, spread through the bites of infected ticks, that is most commonly caused in the United States by infection with the bacterium Ehrlichia chaffeensis. We retrospectively reviewed samples from an 18-month study of ehrlichiosis in the United States and found that E. ewingii was present in 10 (9.2%) of 109 case-patients with ehrlichiosis, a higher rate of infection with this species than had previously been reported. Two patients resided in New Jersey and Indiana, where cases have not been reported. All patients with available case histories recovered. Our study suggests a higher prevalence and wider geographic distribution of E. ewingii in the United States than previous reports have indicated.
Assuntos
Ehrlichia/classificação , Ehrlichiose/diagnóstico , Ehrlichiose/epidemiologia , Adulto , Idoso , Doenças dos Animais/epidemiologia , Animais , Antibacterianos/uso terapêutico , Anticorpos Antibacterianos/imunologia , Ehrlichia/genética , Ehrlichia/imunologia , Ehrlichiose/tratamento farmacológico , Ehrlichiose/microbiologia , Ensaio de Imunoadsorção Enzimática , Humanos , Masculino , Vigilância da População , Prevalência , RNA Ribossômico 16S/genética , Estudos Retrospectivos , Resultado do Tratamento , Estados Unidos/epidemiologia , Adulto JovemRESUMO
Current infectious disease molecular tests are largely pathogen specific, requiring test selection based on the patient's symptoms. For many syndromes caused by a large number of viral, bacterial, or fungal pathogens, such as respiratory tract infections, this necessitates large panels of tests and has limited yield. In contrast, next-generation sequencing-based metagenomics can be used for unbiased detection of any expected or unexpected pathogen. However, barriers for its diagnostic implementation include incomplete understanding of analytical performance and complexity of sequence data analysis. We compared detection of known respiratory virus-positive (n= 42) and unselected (n= 67) pediatric nasopharyngeal swabs using an RNA sequencing (RNA-seq)-based metagenomics approach and Taxonomer, an ultrarapid, interactive, web-based metagenomics data analysis tool, with an FDA-cleared respiratory virus panel (RVP; GenMark eSensor). Untargeted metagenomics detected 86% of known respiratory virus infections, and additional PCR testing confirmed RVP results for only 2 (33%) of the discordant samples. In unselected samples, untargeted metagenomics had excellent agreement with the RVP (93%). In addition, untargeted metagenomics detected an additional 12 viruses that were either not targeted by the RVP or missed due to highly divergent genome sequences. Normalized viral read counts for untargeted metagenomics correlated with viral burden determined by quantitative PCR and showed high intrarun and interrun reproducibility. Partial or full-length viral genome sequences were generated in 86% of RNA-seq-positive samples, allowing assessment of antiviral resistance, strain-level typing, and phylogenetic relatedness. Overall, untargeted metagenomics had high agreement with a sensitive RVP, detected viruses not targeted by the RVP, and yielded epidemiologically and clinically valuable sequence information.
Assuntos
Metagenômica/métodos , Reação em Cadeia da Polimerase/métodos , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Análise de Sequência de RNA/métodos , Vírus/classificação , Vírus/isolamento & purificação , Pré-Escolar , Biologia Computacional/métodos , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Nasofaringe/virologia , Estudos Retrospectivos , Vírus/genéticaRESUMO
The genus Nocardia has undergone rapid taxonomic expansion in recent years, and an increasing number of species are recognized as human pathogens. Many established species have predictable antimicrobial susceptibility profiles, but sufficient information is often not available for recently described organisms. Additionally, the effectiveness of sulfonamides as first-line drugs for Nocardia has recently been questioned. This led us to review antimicrobial susceptibility patterns for a large number of molecularly identified clinical isolates. Susceptibility results were available for 1,299 isolates representing 39 different species or complexes, including 11 that were newly described, during a 6-year study period. All tested isolates were susceptible to linezolid. Resistance to trimethoprim-sulfamethoxazole (TMP-SMX) was rare (2%) except among Nocardia pseudobrasiliensis (31%) strains and strains of the N. transvalensis complex (19%). Imipenem susceptibility varied for N. cyriacigeorgica and N. farcinica, as did ceftriaxone susceptibility of the N. nova complex. Resistance to more than one of the most commonly used drugs (amikacin, ceftriaxone, TMP-SMX, and imipenem) was highest for N. pseudobrasiliensis (100%), N. transvalensis complex (83%), N. farcinica (68%), N. puris (57%), N. brasiliensis (51%), N. aobensis (50%), and N. amikacinitolerans (43%). Thus, while antimicrobial resistance can often be predicted, susceptibility testing should still be considered when combination therapy is warranted, for less well characterized species or those with variable susceptibility profiles, and for patients with TMP-SMX intolerance.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Nocardia/efeitos dos fármacos , Filogenia , Acetamidas/farmacologia , Amicacina/farmacologia , Ceftriaxona/farmacologia , Humanos , Imipenem/farmacologia , Linezolida , Testes de Sensibilidade Microbiana , Nocardia/classificação , Nocardia/genética , Nocardia/isolamento & purificação , Nocardiose/tratamento farmacológico , Nocardiose/microbiologia , Oxazolidinonas/farmacologia , Especificidade da Espécie , Combinação Trimetoprima e Sulfametoxazol/farmacologiaRESUMO
Neonatal Streptococcus agalactiae infections cause significant morbidity and mortality, and antenatal screening is recommended. We compared three U.S. Food and Drug Administration (FDA)-cleared nucleic acid amplification tests (NAATs) to culture using 314 vaginal/rectal swabs after 18 to 24 h (recommended period) and 4 to 8 h (shortened period) of broth enrichment. Agreement of the NAATs with each other was high (97.1% to 98.4%), but culture was less sensitive than all NAATs (67% to 73%). A shortened period of broth culture enrichment resulted in 1 false-negative result in 68 (1.5%). The NAATs performed comparably and were more sensitive than culture.
Assuntos
Técnicas Bacteriológicas/métodos , Programas de Rastreamento/métodos , Técnicas de Diagnóstico Molecular/métodos , Complicações Infecciosas na Gravidez/genética , Diagnóstico Pré-Natal/métodos , Infecções Estreptocócicas/diagnóstico , Streptococcus agalactiae/isolamento & purificação , Erros de Diagnóstico , Feminino , Humanos , Recém-Nascido , Gravidez , Reto/microbiologia , Sensibilidade e Especificidade , Infecções Estreptocócicas/microbiologia , Fatores de Tempo , Estados Unidos , Vagina/microbiologiaRESUMO
The accurate detection and typing of high-risk human papillomavirus (HPV) are critical for cervical cancer screening. The Hybrid Capture 2 (hc2) and cobas HPV tests showed high agreement for cervical samples (94.4%, κ=0.72, n=693) and moderate agreement for vaginal samples (κ=0.62, n=108). The HPV16 and HPV18 results were highly consistent between the cobas and Linear Array tests (κ≥0.96, n=197). Three hc2-negative vaginal samples were repeatedly invalid by the cobas test due to ß-globin control failures, highlighting amplification control benefits. No cross-contamination was detected in a challenge experiment.
Assuntos
Colo do Útero/virologia , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , Vagina/virologia , Adulto , Idoso , DNA Viral/genética , Detecção Precoce de Câncer/métodos , Feminino , Genótipo , Testes de DNA para Papilomavírus Humano , Humanos , Pessoa de Meia-Idade , Esfregaço Vaginal/métodosRESUMO
Pediatric pneumonia can be severe and result in empyema. Next-generation sequencing (NGS) may broadly detect pathogens though, optimal timing and impact of sample type on diagnostic yield is unknown. This is a prospective, single-center pilot study of children aged 3 months through 17 years admitted to the PICU with a primary diagnosis of complicated pneumonia. Plasma, endotracheal, nasopharyngeal, and pleural fluid samples were collected at three time points during hospitalization. After nucleic acid extraction, combined libraries were enriched with an NGS enrichment panel kit (RPIP, Illumina), sequenced and quantitative organism detections were analyzed. NGS identified the same bacterial pathogen as traditional testing in all samples, regardless of antibiotic pre-treatment or time collected. Conventional culture methods only identified the pathogen reliably in invasively obtained pleural fluid or endotracheal aspirates. Future application of NGS may allow for non-invasive pathogen detection at a broader range of time points and more targeted antibiotic coverage.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Criança , Lactente , Pré-Escolar , Estudos Prospectivos , Adolescente , Projetos Piloto , Masculino , Feminino , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/classificação , Pneumonia Bacteriana/diagnóstico , Pneumonia Bacteriana/microbiologia , Nasofaringe/microbiologia , Pneumonia/microbiologia , Pneumonia/diagnósticoRESUMO
Sequencing-based pathogen identification directly from clinical specimens requires time-consuming interpretation, especially with mixed chromatograms when multiple microorganisms are detected. We assessed RipSeq Mixed software for human papillomavirus (HPV) genotyping by comparison to the linear array HPV genotyping assay. RipSeq Mixed provided rapid, sequencing-based HPV typing for single-type infections and coinfections with 2 types.
Assuntos
Coinfecção/virologia , Biologia Computacional/métodos , DNA Viral/química , Papillomaviridae/classificação , Papillomaviridae/genética , Infecções por Papillomavirus/virologia , Análise de Sequência de DNA/métodos , DNA Viral/genética , DNA Viral/isolamento & purificação , Humanos , Papillomaviridae/isolamento & purificação , SoftwareRESUMO
BACKGROUND: The illumigene® (Meridian Bioscience, Inc., Cincinnati, OH) and GeneOhm® (BD Diagnostics, La Jolla, CA) Clostridium difficile assays target the tcdA gene and tcdB gene, respectively. We assessed the use of tcdA as the molecular target in the illumigene® C. difficile loop-mediated amplification assay in detecting a wide variety of C. difficile strains including those with tcdA deletions. METHODS: We tested 38 C. difficile strains and 108 patient stool specimens using the illumigene® assay. The GeneOhm® real-time polymerase chain reaction (PCR) assay served as the reference method. Discordant results were resolved by repeat testing, anaerobic culture, and a laboratory-developed real-time PCR targeting tcdA and tcdB. RESULTS: Both illumigene® and GeneOhm® assays detected all 37 C. difficile toxin B(+) strains representing seven toxinotypes and including four toxin A(-) B(+) isolates. No cross-reactivity with 20 other Clostridium species or toxin-negative C. difficile was observed in either assay. Among patient stool specimens, agreement was 94.4% (102/108). After discordant result resolution, agreement was 96.3% (104/108). Specimens with initially discordant results had target concentrations approaching the limit of detection for the two commercial assays. Discordance appeared unrelated to whether tcdA or tcdB was the amplification target. CONCLUSION: The tcdA 5' region used by the illumigene® assay is a practical target for toxigenic C. difficile detection.
Assuntos
Toxinas Bacterianas/genética , Clostridioides difficile/genética , Enterocolite Pseudomembranosa/microbiologia , Enterotoxinas/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Clostridioides difficile/isolamento & purificação , Enterocolite Pseudomembranosa/diagnóstico , Fezes/microbiologia , Humanos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
BACKGROUND: The SARS-CoV-2 pandemic has underscored the need for field specimen collection and transport to diagnostic and public health laboratories. Self-collected nasal swabs transported without dependency on a cold chain have the potential to remove critical barriers to testing, expand testing capacity, and reduce opportunities for exposure of health professionals in the context of a pandemic. OBJECTIVE: We compared nasal swab collection by study participants from themselves and their children at home to collection by trained research staff. METHODS: Each adult participant collected 1 nasal swab, sampling both nares with the single swab, after which they collected 1 nasal swab from 1 child. After all the participant samples were collected for the household, the research staff member collected a separate single duplicate sample from each individual. Immediately after the sample collection, the adult participants completed a questionnaire about the acceptability of the sampling procedures. Swabs were placed in temperature-stable preservative and respiratory viruses were detected by shotgun RNA sequencing, enabling viral genome analysis. RESULTS: In total, 21 households participated in the study, each with 1 adult and 1 child, yielding 42 individuals with paired samples. Study participants reported that self-collection was acceptable. Agreement between identified respiratory viruses in both swabs by RNA sequencing demonstrated that adequate collection technique was achieved by brief instructions. CONCLUSIONS: Our results support the feasibility of a scalable and convenient means for the identification of respiratory viruses and implementation in pandemic preparedness for novel respiratory pathogens.
RESUMO
Sequencing of the 16S rRNA gene (16S) is a reference method for bacterial identification. Its expanded use has led to increased recognition of novel bacterial species. In most clinical laboratories, novel species are infrequently encountered, and their pathogenic potential is often difficult to assess. We reviewed partial 16S sequences from >26,000 clinical isolates, analyzed during February 2006-June 2010, and identified 673 that have <99% sequence identity with valid reference sequences and are thus possibly novel species. Of these 673 isolates, 111 may represent novel genera (<95% identity). Isolates from 95 novel taxa were recovered from multiple patients, indicating possible clinical relevance. Most repeatedly encountered novel taxa belonged to the genera Nocardia (14 novel taxa, 42 isolates) and Actinomyces (12 novel taxa, 52 isolates). This systematic approach for recognition of novel species with potential diagnostic or therapeutic relevance provides a basis for epidemiologic surveys and improvement of sequence databases and may lead to identification of new clinical entities.
Assuntos
Bactérias/classificação , Bactérias/isolamento & purificação , Técnicas de Tipagem Bacteriana/métodos , Bactérias/genética , Humanos , Anotação de Sequência Molecular , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
No clinical isolates have been reported for the recently described thermoactinomycete Kroppenstedtia eburnea. Between 2006 and 2011, we obtained 14 clinical isolates from patients in 9 U.S. states. Here we report growth characteristics, 16S rRNA gene sequencing, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry-based identification, and antimicrobial susceptibility profiles of this recently described organism.
Assuntos
Bacillales/isolamento & purificação , Infecções por Bactérias Gram-Positivas/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/farmacologia , Bacillales/química , Bacillales/genética , Bacillales/crescimento & desenvolvimento , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Feminino , Humanos , Lactente , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Estados UnidosRESUMO
Chronic fatigue syndrome (CFS) is a multisystem disorder characterized by prolonged and severe fatigue that is not relieved by rest. Attempts to treat CFS have been largely ineffective primarily because the etiology of the disorder is unknown. Recently, CFS has been associated with xenotropic murine leukemia virus-related virus (XMRV) as well as other murine leukemia virus (MLV)-related viruses, though not all studies have found these associations. We collected blood samples from 100 CFS patients and 200 self-reported healthy volunteers from the same geographical area. We analyzed these in a blind manner using molecular, serological, and viral replication assays. We also analyzed samples from patients in the original study that reported XMRV in CFS patients. We did not find XMRV or related MLVs either as viral sequences or infectious viruses, nor did we find antibodies to these viruses in any of the patient samples, including those from the original study. We show that at least some of the discrepancy with previous studies is due to the presence of trace amounts of mouse DNA in the Taq polymerase enzymes used in these previous studies. Our findings do not support an association between CFS and MLV-related viruses, including XMRV, and the off-label use of antiretrovirals for the treatment of CFS does not seem justified at present.