3-氨基苯硼酸联合乙二胺四乙酸碳青霉烯酶抑制增强试验检测肠杆菌目细菌产碳青霉烯酶的研究

目的 了解3-氨基苯硼酸(PBA)联合乙二胺四乙酸(EDTA)碳青霉烯酶抑制剂增强试验(PBA-EDTA法)用于检测肠杆菌目细菌产碳青霉烯酶的检测结果.方法 使用PBA-EDTA法测定275株经金标准PCR及DNA测序已明确为产碳青霉烯酶肠杆菌目细菌产碳青霉烯酶结果的符合率,并与CLSI推荐的mCIM联合eCIM改良碳...     

Clinical evaluation of the rapid nucleic acid amplification point-of-care test (Smart Gene SARS-CoV-2) in the analysis of nasopharyngeal and anterior nasal samples

IntroductionSmart Gene is a point-of-care (POC)-type automated molecular testing platform that can be performed with 1 min of hands-on-time. Smart Gene SARS-CoV-2 is a newly developed Smart Gene molecular assay for the detection of SARS-CoV-2. The analytical and clinical performance of Smart Gene SARS-CoV-2 has not been evaluated.MethodsNasopharyngeal and anterior nasal samples were prospectively collected from subjects referred to the local PCR center from March 25 to July 5, 2021. Two swabs were simultaneously obtained for the Smart Gene SARS-CoV-2 assay and the reference real-time RT-PCR assay, and the results of Smart Gene SARS-CoV-2 were compared to the reference real-time RT-PCR assay.ResultsAmong a total of 1150 samples, 68 of 791 nasopharyngeal samples and 51 of 359 anterior nasal samples were positive for SARS-CoV-2 in the reference real-time RT-PCR assay. In the testing of nasopharyngeal samples, Smart Gene SARS-CoV-2 showed the total, positive and negative concordance of 99.2% (95% confidence interval [CI]: 98.4–99.7%), 94.1% (95% CI: 85.6–98.4%) and 99.7% (95% CI: 99.0–100%), respectively. For anterior nasal samples, Smart Gene SARS-CoV-2 showed the total, positive and negative concordance of 98.9% (95% CI: 97.2–99.7%), 98.0% (95% CI: 89.6–100%) and 99.0% (95% CI: 97.2–99.8%), respectively. In total, 5 samples were positive in the reference real-time RT-PCR assay and negative in the Smart Gene SARS-CoV-2 assay, whereas 5 samples were negative in the reference real-time RT-PCR assay and positive in the Smart Gene SARS-CoV-2 assay.ConclusionSmart Gene SARS-CoV-2 showed sufficient analytical performance for the detection of SARS-CoV-2 in nasopharyngeal and anterior nasal samples.     

High clinical performance and quantitative assessment of antibody kinetics using a dual recognition assay for the detection of SARS-CoV-2 IgM and IgG antibodies

ObjectivesSeveral serological SARS-CoV-2 immunoassays have been developed recently but require external validation before widespread use. This study aims at assessing the analytical and clinical performance of the iFlash® anti-SARS-CoV-2 chemiluminescence assay for the detection of both IgM and IgG antibodies. The kinetics of the antibody response was also evaluated.Design & MethodsThe precision, carry-over, linearity, limit of blank, detection and quantification were assessed. Sensitivity analysis was performed by using 178 sera collected from 154 RT-PCR confirmed COVID-19 patients. The specificity analysis was performed from 75 selected non-SARS-CoV-2 sera with a potential cross-reaction to the SARS-CoV-2 immunoassay.ResultsThis iFlash® SARS-CoV-2 assay showed excellent analytical performance. After 2 weeks since symptom onset, the sensitivities for IgM and IgG were 62.2% (95% CI: 52.3–71.2%) and 92.9%% (95% CI: 85.7–96.7%), respectively by using the cut-off provided by the manufacturer. After cut-off optimization (i.e. >2.81 for IgM and >4.86 for IgG), the sensitivity for IgM and IgG were 81.6 (95% CI: 72.7–88.1%) and 95.9% (95% CI: 89.4–98.7%), respectively. Optimized cut-off for IgG improved the sensitivity to reach 100% (95%CI: 87.6–100) from 28 days since symptom onset.ConclusionsThis study shows that the iFlash® SARS-CoV-2 assay from YHLO biotechnology, has satisfactory analytical performance. Nevertheless, the sensitivity of the IgM is limited for a proper clinical use compared to IgG. The determination of anti-SARS-CoV-2 IgG antibodies from 28 days since symptom onset was associated with high sensitivity, especially using optimized cut-offs (i.e. 100%).     

Comparison of the performance of three carbapenem inactivation methods for the detection of carbapenemase-producing gram-negative bacilli

IntroductionThe carbapenem inactivation method test (CIM) was developed as a method for detecting carbapenemase-producing Gram-negative bacilli, and the modified CIM (mCIM) was recommended by the CLSI for as an improved method in M100-S27. However, few studies have evaluated the influence of bacterial species and genotype on its sensitivity and specificity. In this study, we evaluate the performance of these improved modified CIM methods with mCIM.MethodsAs strains, clinical isolates from Naga Municipal Hospital and stored strains from the Study of Bacterial Resistance in the Kinki Region of Japan were used. The mCIM, CIM-Tris, and simple CIM (sCIM) test methods were applied to 120 Enterobacterales, 40 Pseudomonas aeruginosa, and 37 Acinetobacter spp. The procedure and criteria for each method were based on the original papers and the CLSI M − 100 S27 documents.ResultsThe sensitivity of the test methods in the detection of carbapenemase in Enterobacterales, Pseudomonas spp., and Acinetobacter spp. was as follows: mCIM, 98.9%, 90.0%, and 76.5%, respectively; CIM-Tris, 94.4%, 100%, 100%; and sCIM 98.9%, 85.0%, 76.5%. All methods showed 100% specificity in Enterobacterales, Pseudomonas spp., and Acinetobacter spp. Each method performed well in the detection of metallo β-lactamase-producing strains, however, the sensitivity tended to be low in the detection of the organisms producing serine-type carbapenemase, such as GES, OXA-23, and OXA-51.ConclusionsCare must be taken when selecting test methods because the sensitivity of the detection differs depending on the bacterial species and genotype.     

Modified Carba NP Test: Simple and rapid method to differentiate KPC‐ and MBL‐producing Klebsiella species

Background

The aim of this study was to evaluate the modified Carba NP test to differentiate KPC (Klebsiella pneumoniae carbapenemase)‐ and MBL (metallo‐β‐lactamase)‐producing Klebsiella species.

Methods

A total of 508 non‐duplicate clinical isolates of Klebsiella spp. were processed by modified Carba NP and combined disc tests which were further confirmed by conventional polymerase chain reaction (PCR), a gold standard method for statistical analysis.

Results

Modified Carba NP test demonstrated 91.7% sensitivity, 100% specificity, 100% positive predictive value (PPV) and 99.8% negative predictive value (NPV) for KPC and 96.7%, 100%, 100%, and 99.5% for MBL detection, respectively.

Conclusion

The performance of modified Carba NP test was significantly better than combined disc test, fulfilling the requirement of simple and rapid test for clinical applications.

     

Epidemiology and molecular characterization of fecal carriage of third-generation cephalosporin-resistant Enterobacterales among elderly residents in Japan

IntroductionThe spread of third-generation cephalosporin-resistant Gram-negative bacteria is a serious concern in acute and post-acute care settings. This study aimed to understand the epidemiology and molecular background of fecal colonization of resistant Enterobacterales in elderly people.MethodsIn December 2015–December 2017, stool or rectal swab samples were collected from 101 elderly patients receiving home care, using long-term care facilities (LTCF), and living in nursing homes repeatedly at 3?9-month intervals. Patient clinical background data were collected from medical records. After phenotypic screening for extended-spectrum β-lactamase (ESBL), AmpC-type β-lactamase or carbapenemase production, drug resistance genes of isolates were analyzed using polymerase chain reaction (PCR). ESBL-producing Escherichia coli isolates obtained from the same patients in repetitive screenings were analyzed using PCR-based ORF typing. Risk factors for persistent carriage of resistant Enterobacterales were analyzed using multivariate analysis.ResultsResistant Enterobacterales isolates were detected in 37 of 101 (36.6%) and 29 of 80 (36.3%) residents in first and second screenings, respectively. ESBL-producing E. coli accounted for 80% isolates, the most common being CTX-M-9-group β-lactamase producers. Molecular epidemiological analysis revealed probable transmissions of ESBL-producing E. coli; 58% of ESBL-producing E. coli colonizers were persistent colonizers at least after 3 -month intervals. Age > 87 years and LTCF residence were independent risk factors for persistent carriage of ESBL-producing E. coli.ConclusionsWe showed, for the first time, high persistent colonization rate of ESBL-producing E. coli among elderly people in post-acute care settings with probable horizontal transmission. We also identified significant risk factors for persistent colonization.     

Sustained postprandial decrease in plasma levels of LDL cholesterol in patients with type‐2 diabetes mellitus

Objective. Low density lipoprotein cholesterol (LDL‐C) is an independent and modifiable risk factor for development of cardiovascular disease (CVD). Postprandial lipid metabolism has been linked to CVD, but little is known about the postprandial LDL‐C profile in patients with type‐2 diabetes (T2DM). We aimed to study the postprandial levels of LDL‐C in T2DM patients. Material and methods. After an overnight fast, 74 T2DM patients, mean age approximately 60 years, were served a standard fat‐rich meal of 3,515 kJ containing 54?% fat, 13?% protein and 33?% carbohydrates. Only drinking water was allowed postprandially. Blood samples were drawn at times 0 (fasting), 1.5, 3.0, 4.5 and 6.0?h (postprandial). In all samples, LDL‐C was measured with modified beta quantification (separation by ultracentrifugation followed by measurement of infranate high density lipoprotein cholesterol (HLD‐C) using a homogeneous assay). Results. At all postprandial times, levels of LDL‐C showed highly significant (p<0.005) decreases compared to time 0 (mean [95?% CI] maximum change in LDL‐C levels at 3.0?h: ?0.16?mmol/L [?0.12; ?0.20]; p<0.001). Independently of fasting LDL‐C levels and ongoing statin therapy, LDL‐C decreased significantly more in female compared to male patients postprandially (mean [95?% CI] maximum unadjusted change versus time 0 in LDL‐C for men [n = 56] at 3.0?h: ?0.14?mmol/L [?0.19; ?0.10], p<0.001; for women [n = 18] at 4.5?h: ?0.26?mmol/L [?0.35; ?0.18], p<0.001; ?0.14?mmol/L [?0.24; ?0.05], p = 0.005 between genders for the mean [95?% CI] fasting adjusted difference at 4.5?h in the change versus time 0 in LDL‐C; gender by time interaction: p = 0.007 (repeated measures mixed model)). Conclusions. In T2DM patients served a fat‐rich meal, levels of LDL‐C decreased significantly more in women compared to men postprandially, irrespective of fasting levels or ongoing statin therapy. This might have implications in the atherosclerotic process and on any difference in the risk of CVD between genders.     

Evaluation of the modified carbapenem inactivation method for the detection of carbapenemase-producing Enterobacteriaceae

Carbapenemase-producing Enterobacteriaceae (CPE) are increasing worldwide. Rapid and accurate detection of CPE is necessary for appropriate antimicrobial treatment and hospital infection control. However, CPE contains some strains that are difficult to detect depending on genotype and MIC value of carbapenem, and a detection method has not been established. The recently reported modified carbapenem inactivation method (mCIM) has been developed in CLSI M100-S27 as a phenotypic technique for detecting carbapenemase activity. In the present study, we examined mCIM as a new CPE detection method using 207 Enterobacteriaceae isolates in comparison with the three existing screening methods of modified Hodge test, Carba NP test and carbapenem inactivation method and evaluated its performance. Consequently, both the sensitivity and specificity of mCIM were 100%, indicating better results than the conventional screening methods. The mCIM is a useful tool for microbiology laboratories due to its simplicity, clear criteria, cost-effectiveness and availability at any laboratory.     

Investigation of age-adjusted D-dimer using an uncommon assay

BackgroundUse of an age-adjusted D-dimer for the evaluation of acute pulmonary embolus (PE) has been prospectively validated in the literature and has become a practice recommendation from major medical societies. Most research on this subject involves the most common D-dimer assays reporting in Fibrinogen Equivalent Units (FEU) with a non-age-adjusted manufacturer-recommended cutoff of 500 ng/ml FEU. Limited research to date has evaluated age-adjustment in assays that report in D-Dimer Units (D-DU), which use a manufacturer-recommended cutoff of 230 ng/ml D-DU. Despite scant evidence, an age-adjusted formula using D-DU has been recently endorsed by the American College of Emergency Physicians (ACEP). This formula seems arbitrary in its derivation and unnecessarily deviates from existing thresholds, thus prompting the creation of our novel-age adjustment formula. The goal of this study was to retrospectively evaluate the test characteristics of our novel age-adjusted D-dimer formula using the D-DU assay in comparison to existing traditional and age-adjusted D-dimer thresholds for the evaluation of acute PE in the ED.MethodsThis was a retrospective chart review at an academic quaternary health system with three EDs and 195,000 combined annual ED visits. Only patients with D-dimer testing and CT PE protocol (CTPE) imaging were included. Admission and discharge diagnosis codes were used to identify acute PE. Outcome measures were sensitivity, specificity, negative predictive value (NPV) and positive predictive value (PPV) of an unadjusted traditional threshold (230) compared with both novel and ACEP-endorsed age adjusted thresholds, (Age × 5) ? 20 and Age × 5 if >50, respectively. Estimates with their exact 95% threshold were performed.Results4846 adult patients were evaluated from January 2012 to July 2017. Group characteristics include a mean age of 52 and a frequency of acute PE diagnosis by CTPE of 8.25%. Traditional D-dimer cutoff demonstrated a sensitivity of 99.8% (95% CI 98.6–100), specificity of 16.7% (95% CI 15.6–17.8) and NPV of 99.9% (95% CI 99.3–100). Our novel age-adjusted D-dimer thresholds had a sensitivity of 97.0% (95% CI 94.8–98.4), specificity of 27.9% (95% CI 26.6–29.2) and NPV of 99.0% (95% CI 98.3–99.5) with the ACEP-endorsed formula demonstrating similar test characteristics.ConclusionUse of an age-adjusted D-dimer on appropriately selected patients being evaluated for acute PE in the ED with a D-DU assay increases specificity while maintaining a high sensitivity and NPV. Both our novel formula and the ACEP-endorsed age-adjusted formula performed well, with our novel formula showing a trend towards improved testing characteristics.     

Comparing three different phenotypic methods for accurate detection of carbapenemase-producing Enterobacterales

BackgroundEarly identification of carbapenemase-producing Enterobacterales (CPE) is highly essential to prevent their dissemination within health care settings.ObjectiveThis study aimed to compare 3 reported phenotypic assays for detecting carbapenemase-producing Enterobacterales (CPE).Methods151 Enterobacterales isolates were collected, the sensitivity and specificity of each test was determined, with molecular genotype serving as the gold standard. The phenotypic evaluations were performed using EDTA-synergistic carbapenem inactivation method (esCIM), EDTA-carbapenem inactivation method (eCIM), and enzyme inhibitor enhancement experiment (EIE).ResultsThe concordance rate was 98% for the EIE for the detection of KPC producer, and 100% for the esCIM and eCIM. Sensitivity differed among the 3 methods, and all assays had excellent sensitivity exceeding 90% for detecting metallo-β-lactamases (MBLs). The specificity of the eCIM, esCIM and EIE was 100%, 100% and 95%. Both eCIM and esCIM were unsatisfactory in detecting multi-enzyme strains (MBL and class A serine carbapenemase) (0/6). However, EIE increased the positive number to six (6/6).ConclusionsThe eCIM, esCIM and EIE can be used to accurately detect and distinguish carbapenemase and is suitable for routine use in most clinical microbiology laboratories.     

The effect of prehabilitation on the range of motion and functional outcomes in patients following the total knee or hip arthroplasty: A pilot randomized trial

Objective: The study investigated the effect of prehabilitation on the quality of life and function in patients having total knee replacement (TKR)/total hip replacement (THR). Methods: A pilot randomized controlled trial with concealed allocation, assessor blinding, and intention-to-treat analysis was conducted. Sixty-four people undergoing elective lower-limb arthroplasty were included. Prehabilitation included one-hour twice-weekly sessions for at least three and a maximum of four weeks prior to surgery. Control participants did not complete any pre-surgical programs. Health utility and quality of life as measured by the EQ-5D-3L and the patient-specific functional scale were the primary outcomes measured before allocation and eight weeks post-operatively. Results: No between-group differences were evident in health utility (main effect of the group ?0.04 (95% Confidence Interval [CI] ?0.16 to 0.08, p = 0.50) or patient-specific functional scale (main effect of the group ?0.59 (95% CI ?1.8 to 0.6, p = 0.73), but the group-by-joint interaction effects for the timed up and go (TUG) (7.6 (95% CI ?0.9 to 16.1, p = 0.08)) and the EQ-5D VAS (?18.3 (95% CI ?41.1 to 4.5), p = 0.11) were larger. Prehabilitation participants’ knee flexion improved by 12.6 degrees (95% CI 5.2–20, p = 0.001). Conclusions: Prehabilitation improved knee flexion, but this did not translate into improved functional mobility or quality of life.     

mCIM 和碳青霉烯酶抑制剂增强试验检测CRE 和CRPA 产酶表型的方法学评价

目的 综合评估改良碳青霉烯酶灭活试验（mCIM）和碳青霉烯酶抑制剂增强试验对碳青霉烯酶检测和分型能力。方法 采用mCIM 和碳青霉烯酶抑制剂增强试验检测中山大学肿瘤防治中心2019 ～ 2022 年临床分离并保存的47 株耐碳青霉烯类肠杆菌目细菌（carbapenem-resistant Enterobacterales, CRE）和42 株耐碳青霉烯类铜绿假单胞菌（carbapenem-resistant Pseudomonas aeruginosa, CRPA）的产酶表型,胶体金法对检测结果进行验证比对,通过卡方检验进行统计分析,并从多角度综合评价。结果 mCIM 检测CRE 和CRPA 的阳性率分别为70.2% 和35.7%,碳青霉烯酶不确定占比为25.5% 和11.9%。增强试验检测47 株CRE:44.7% 产B 类金属β 内酰胺酶,17.0% 产A 类碳青霉烯酶,不产A 或B 类碳青霉烯酶的菌株占38.3%;增强试验检测42 株CRPA:2.4% 产B 类金属β 内酰胺酶,90.4% 产A 类碳青霉烯酶,同时产A 类碳青霉烯酶和B 类金属β- 内酰胺酶的菌株占4.8%,不产A 或B 类碳青霉烯酶的菌株占2.4%。两种方法检测CRE 差异有统计学意义（χ2=17.803,P =0.01）,检测CRPA 差异无统计学意义（χ2=4.632,P=0.592）。胶体金法验证:产碳青霉烯酶的阳性符合率为84.6%,产丝氨酸酶的阳性符合率为80%,产金属酶的阳性符合率为100%。结论 检测CRE两种方法均适用且差异不大, CRPA更推荐碳青霉烯酶抑制剂增强试验,胶体金值得推广,临床实验室应根据实际条件选择检测方法。     

Field evaluation of a dual rapid Human Immunodeficiency Virus and treponemal syphilis rapid test in community-based clinics in Los Angeles and New York

Introduction

Dual human immunodeficiency virus/syphilis rapid diagnostic devices can play an important role in prevention efforts. The field performance of the INSTI Multiplex HIV-1/HIV-2/Syphilis Antibody Test (Multiplex) was evaluated.

Diagnostic performance of a novel digital immunoassay (RapidTesta SARS-CoV-2): A prospective observational study with nasopharyngeal samples

IntroductionDigital immunoassays are generally regarded as superior tests for the detection of infectious disease pathogens, but there have been insufficient data concerning SARS-CoV-2 immunoassays.MethodsWe prospectively evaluated a novel digital immunoassay (RapidTesta SARS-CoV-2). Two nasopharyngeal samples were simultaneously collected for antigen tests and Real-time RT-PCR.ResultsDuring the study period, 1127 nasopharyngeal samples (symptomatic patients: 802, asymptomatic patients: 325) were evaluated. For digital immunoassay antigen tests, the sensitivity was 78.3% (95% CI: 67.3%–87.1%) and the specificity was 97.6% (95% CI: 96.5%–98.5%). When technicians visually analyzed the antigen test results, the sensitivity was 71.6% (95% CI: 59.9%–81.5%) and the specificity was 99.2% (95% CI: 98.5%–99.7%). Among symptomatic patients, the sensitivity was 89.4% (95% CI; 76.9%–96.5%) with digital immunoassay antigen tests, and 85.1% (95% CI; 71.7%–93.8%) with visually analyzed the antigen test, respectively.ConclusionsThe sensitivity of digital immunoassay antigen tests was superior to that of visually analyzed antigen tests, but the rate of false-positive results increased with the introduction of a digital immunoassay device.     

Longitudinal associations of sauna bathing with inflammation and oxidative stress: the KIHD prospective cohort study

Purpose: We sought to determine cross-sectional and longitudinal associations of frequency of sauna bathing with high sensitivity C-reactive protein (hsCRP), fibrinogen, leucocyte count and gamma-glutamyltransferase (GGT).

Design: Baseline sauna bathing habits were assessed in 2269 men aged 42–61 years. Concentrations of hsCRP, fibrinogen, leucocyte count, and GGT were determined at baseline and 11 years later. The associations of sauna bathing frequency with baseline and 11-year hsCRP, fibrinogen, leucocyte count, and GGT levels were examined using robust multivariate regression analyses.

Results: In baseline analysis, 4–7 sauna sessions/week (compared with 1 sauna session/week) was associated with ?0.84?mg/l (95% CI, ?1.55, ?0.14; p?=?.019) lower hsCRP; ?0.07?g/l (95% CI, ?0.15, 0.02; p?=?.112) lower fibrinogen; and ?0.28?×?10⁹/l (95% CI, ?0.51, ?0.06; p?=?.015) lower leucocyte count, after multivariable adjustment. In longitudinal analysis, the corresponding estimates were ?1.66?mg/l (95% CI, ?3.13, ?0.19; p?=?.027); ?0.16?g/l (95% CI, ?0.31, ?0.02; p?=?.031); and ?0.49?×?10⁹/l (95% CI, ?0.85, ?0.14; p?=?.007) respectively. Sauna bathing frequency was not associated with GGT at baseline and 11 years.

Conclusion: Observational evidence supports the hypothesis that reduction in inflammation may be one of the pathways linking frequent sauna bathing with decreased risk of acute and chronic disease conditions.

• KEY MESSAGES

• Cross-sectional evidence or short-term studies suggest Finnish sauna bathing may exert its beneficial health effects via reduction in inflammation and oxidative stress; however, the long-term effects of sauna bathing on these outcomes are uncertain.

• In this population-based prospective cohort study, frequent sauna sessions significantly decreased levels of inflammatory markers at baseline and 11-year follow-up; but had no effect on oxidative stress.

• The health benefits of sauna bathing may in part be mediated via reduced systemic inflammation.

     

Evaluation of the BinaxNOW PBP2a assay for the direct detection of methicillin resistance in Staphylococcus aureus from positive blood culture bottles

BinaxNOW® PBP2a rapid immunochromatographic assay is a novel test for the identification of methicillin resistance in Staphylococcus aureus from clinical blood culture samples based on detection of penicillin binding protein 2a. We have evaluated the utility of this assay to do a rapid diagnostic of methicillin susceptibility directly from blood culture bottles after identification of S. aureus in positive bottles by matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry. Twenty of 21 methicillin-resistant S. aureus (MRSA) samples from blood cultures were positive on direct immunochromatographic testing (sensitivity 95.24%, 95% confidence interval [CI] 74.13% to 99.75%), whereas 37 methicillin-susceptible S. aureus (MSSA) samples were negative (specificity 100%, 95% CI 88.99% to 99.75%). The combined use of MALDI-TOF mass spectrometry and BinaxNOW® PBP2a test is useful for the rapid identification of both MRSA and MSSA from blood cultures.