Intraindividual Scale Comparison in Clinical Diagnostic Methods: A Review of Elementary Methods

This text tries to summarize proposals on the statistical comparison of clinical measurement methods based on bivariate normally distributed outcome. It mainly concentrates on design adequate intraindividual comparison of variances in repeated measurements and reviews univariate graphical and numerical representations of corresponding deviations. The latter will be illustrated by the comparison of two cardiac imaging procedures in 28 probands.     

Calreticulin Mutations in Myeloproliferative Neoplasms: Comparison of Three Diagnostic Methods

Calreticulin (CALR) mutations have recently been reported in 70–84% of JAK2V617F-negative myeloproliferative neoplasms (MPN), and this detection has become necessary to improve the diagnosis of MPN. In a large single-centre cohort of 298 patients suffering from Essential Thrombocythemia (ET), the JAK2V617F, CALR and MPL mutations were noted in 179 (60%), 56 (18.5%) and 13 (4.5%) respectively. For the detection of the CALR mutations, three methods were compared in parallel: high-resolution melting-curve analysis (HRM), product-sizing analysis and Sanger sequencing. The sensitivity for the HRM, product-sizing analysis and Sanger sequencing was 96.4%, 98.2% and 89.3% respectively, whereas the specificity was 96.3%, 100% and 100%. In our cohort, the product-sizing analysis was the most sensitive method and was the easiest to interpret, while the HRM was sometimes difficult to interpret. In contrast, when large series of samples were tested, HRM provided results more quickly than did the other methods, which required more time. Finally, the sequencing method, which is the reference method, had the lowest sensitivity but can be used to describe the type of mutation precisely. Altogether, our results suggest that in routine laboratory practice, product-sizing analysis is globally similar to HRM for the detection of CALR mutations, and that both may be used as first-line screening tests. If the results are positive, Sanger sequencing can be used to confirm the mutation and to determine its type. Product-sizing analysis provides sensitive and specific results, moreover, with the quantitative measurement of CALR, which might be useful to monitor specific treatments.     

结直肠癌诊断方法的应用与评价

结直肠癌(colorectal cancer,CRC)是人类常见肿瘤之一,早期诊断对预防CRC的发病显得尤为重要.随着医学技术的发展,CRC的常规诊断方法已取得了很大进展,目前用于诊断CRC的检查方法主要有粪便排泄物检查、结肠镜检查、CT结肠成像检查、肿瘤分子标记物等,然而各诊断方法的敏感性和特异性存在一定差异.本文主要对目前CRC的常见诊断方法进行分析和评价,以期为CRC的临床预防和早期诊断提供更多的理论依据.     

啮齿类螺杆菌不同检测方法的比较

目的确定用于实验啮齿类动物常规螺杆菌检测方法。方法选用针对螺杆菌属16S rRNA属特异性的4对引物进行了引物选择、取材部位以及与分离培养法比较。结果引物P7/P8的敏感性优于其它3对,检出限可达0.01 pg,对阳性动物群检出率80%-100%;分离培养法检出率40%-50%;盲肠、结肠检出率无显著差异。结论以引物P7/P8的PCR方法作为啮齿类实验动物螺杆菌初筛方法,分离培养法作为验证方法,取材部位可在盲肠或结肠。     

White-Nose Syndrome Disease Severity and a Comparison of Diagnostic Methods

White-nose syndrome is caused by the fungus Pseudogymnoascus destructans and has killed millions of hibernating bats in North America but the pathophysiology of the disease remains poorly understood. Our objectives were to (1) assess non-destructive diagnostic methods for P. destructans infection compared to histopathology, the current gold-standard, and (2) to evaluate potential metrics of disease severity. We used data from three captive inoculation experiments involving 181 little brown bats (Myotis lucifugus) to compare histopathology, quantitative PCR (qPCR), and ultraviolet fluorescence as diagnostic methods of P. destructans infection. To assess disease severity, we considered two histology metrics (wing area with fungal hyphae, area of dermal necrosis), P. destructans fungal load (qPCR), ultraviolet fluorescence, and blood chemistry (hematocrit, sodium, glucose, pCO₂, and bicarbonate). Quantitative PCR was most effective for early detection of P. destructans, while all three methods were comparable in severe infections. Correlations among hyphae and necrosis scores, qPCR, ultraviolet fluorescence, blood chemistry, and hibernation duration indicate a multi-stage pattern of disease. Disruptions of homeostasis occurred rapidly in late hibernation. Our results provide valuable information about the use of non-destructive techniques for monitoring, and provide novel insight into the pathophysiology of white-nose syndrome, with implications for developing and implementing potential mitigation strategies.     

九种单核细胞增生性李斯特菌检测技术效果比较及评价

采用传统生物学方法、显色培养基方法、自制三抗体夹心ELISA试剂盒(TAS-ELISA)、基因组直接PCR(DNA Direct-PCR)、菌裂解直接PCR(Bacterium Direct-PCR)、免疫捕捉PCR(IC-PCR)、生物PCR(Bio-PCR)、SYBR Green染料实时荧光PCR(SYBR Green Real time-PCR)、探针实时荧光PCR(TaqMan Real time-PCR)检测纯菌液及模拟带菌食品中的单核细胞增生性李斯特菌(Listeria monocytogenes,LM)。结果表明:传统生物学培养方法出现假阳性;TAS-ELISA、DNADirect-PCR、Bacterium Direct-PCR、IC-PCR最低检测限分别为106、102、105、104CFU/ml;显色培养基、SYBR Green Real time-PCR灵敏度达102CFU/ml;Bio-PCR、TaqMan Real time-PCR检测灵敏度最高,均为101CFU/ml。显色培养基、TAS-ELISA操作简单,适合大量样品的初检;IC-PCR具有灵敏、特异、快速、经济的优点,特别适合大体积样品中微量病原的检测;Bio-PCR、TaqMan Real time-PCR灵敏度高、特异性好,适合阳性样品的复检以及科研使用。     

Onychomycosis in Malaysia

The common etiological agents of onychomycosis are dermatophytes, molds and yeasts. A mycological nail investigation of onychomycosis using direct microscopy and culture was conducted by the Mycology Unit, Department of Medical Microbiology, University of Malaya from March 1996 to November 1998. The study involved 878 nail clippings or subungal scrapings from subjects with onychomycosis. On direct microcopy examination, 50% of the specimens were negative for fungal elements. On culture, 373 specimens had no growth; bacteria were isolated from 15 nail specimens. Among the 490 specimens with positive fungal cultures, 177 (36.1%) were dermatophytes, 173 (35.5%) were molds and 130 (26.5%) were Candida. There were 2% (10/490) mixed infections of molds, yeasts and dermatophytes. Trichophyton rubrum (115/177) and Trichophyton mentagrophytes (59/177) were the main dermatophytes isolated. The molds isolated were predominantly Aspergillus niger (61/173), Aspergillus nidulans (30/173), Hendersonula toruloidea (26/173) and Fusarium species (16/173). 96.9% of the Candida species identified were Candida albicans.This revised version was published online in October 2005 with corrections to the Cover Date.     

Prevalence and Comparison of Diagnostic Methods for Trichomonas vaginalis Infection in Pregnant Women in Argentina

The objectives of this study were to conduct a prevalence survey of trichomoniasis in pregnant women and to evaluate the utility of different methods for its diagnosis. A total of 597 vaginal exudates from pregnant women who were examined at the Hospital de Clinicas in Buenos Aires, Argentina from 1 August 2005 to 31 January 2007, were prospectively and consecutively evaluated. The investigation of Trichomonas vaginalis was made by different microscopic examinations, and culture on liquid medium. The sensitivity and specificity of the microscopic examinations were assessed considering culture on liquid medium as the "gold standard". The prevalence of T. vaginalis obtained by culture on liquid medium was 4.0% (24/597). The prevalence of T. vaginalis obtained by direct wet smear, prolonged May-Grunwald Giemsa staining, and sodium acetate-formalin (SAF)/methylene blue staining-fixing technique was 1.8%, 2.3% and 2.5%, respectively. The sensitivity of the direct wet smear was 45.8%, that of the prolonged May-Grunwald Giemsa staining was 58.3%, and that of the SAF/methylene blue method was 62.5%. Considering the 3 microscopic examinations altogether, the sensitivity rose to 66.7% and the specificity was 100% for all of them. This is the first time that the prevalence data of T. vaginalis by culture in pregnant women are published in Argentina. Due to the low sensitivity obtained by microscopy in asymptomatic pregnant women, the use of the liquid medium is recommended during pregnancy, in order to provide an early diagnosis and treatment.     

An Evaluation of Methods for the Estimation of Sensitivity and Specificity of Site-Specific Diagnostic Tests

The performance of diagnostic tests is often evaluated by estimating their sensitivity and specificity with respect to a traditionally accepted standard test regarded as a “gold standard” in making the diagnosis. Correlated samples of binary data arise in many fields of application. The fundamental unit for analysis is occasionally the site rather than the subject in site-specific studies. Statistical methods that take into account the within-subject corelation should be employed to estimate the sensitivity and the specificity of diagnostic tests since site-specific results within a subject can be highly correlated. I introduce several statistical methods for the estimation of the sensitivity and the specificity of sitespecific diagnostic tests. I apply these techniques to the data from a study involving an enzymatic diagnostic test to motivate and illustrate the estimation of the sensitivity and the specificity of periodontal diagnostic tests. I present results from a simulation study for the estimation of diagnostic sensitivity when the data are correlated within subjects. Through a simulation study, I compare the performance of the binomial estimator pCBE, the ratio estimator pCBE, the weighted estimator pCWE, the intracluster correlation estimator pCIC, and the generalized estimating equation (GEE) estimator PCGEE in terms of biases, observed variances, mean squared errors (MSE), relative efficiencies of their variances and 95 per cent coverage proportions. I recommend using PCBE when σ == 0. I recommend use of the weighted estimator PCWE when σ = 0.6. When σ == 0.2 or σ == 0.4, and the number of subjects is at least 30, PCGEE performs well.     

A Comparison of Cytochemical Methods for the Rapid Evaluation of Microalgal Viability

We compared various rapid methods for the evaluation of cell viability of 30 algal strains from 15 genera using dyes for both light and fluorescence microscopy. Algal strains demonstrated considerable staining specificity. Staining with fluorescein diacetate helps distinguish between living and dead cells and also predicts the physiological state of the unicellular alga.     

Comparison and Evaluation of Three Methods of Estimation of the Johnson Sb Distribution

Thre methods of estimating the parameters of the Johnson S₆ distribution were tested by simulation. The maximum likelihood method, the method based on percentiles of a sample and the method based on moments of a transformed random variable were taken into consideration. Many sets of samples were generated differing in sizes and in the actual values of parameters, whereupon the parameters were estimated by the three methods. It was proved that if the sample is small or the skewness of the distribution is considerable, the maximum likelihood estimates can assume preposterous values. The method based on moments is recommended due to its simplicity and to the fact that the estimates, though usually biased, never assume absurd values.     

脑动静脉畸形的MRA诊断技术及诊断价值

目的：回顾性分析脑动静脉畸形（AVM）的MRA诊断方法厦诊断价值。方法：51例AVM病人,男30人,女21人,年龄为9—66岁。均进行MR平扫压3D—TOFMBA或3D—PCMBA扫描。结果：删能清晰显示瘤巢压AVM的继发改变,MBA可基本显示AVM的全貌。结论：船认是诊断AVM的有效方法,但常需结合DSA。     

脑动静脉畸形的MRA诊断技术及诊断价值

目的:回顾性分析脑动静脉畸形(AVM)的MRA诊断方法及诊断价值。方法:51例AVM病人,男30人,女21人,年龄为9-66岁。均进行MR平扫及3D-TOF MRA或3D-PC MRA扫描。结果:MRI能清晰显示瘤巢及AVM的继发改变,MRA可基本显示AVM的全貌。结论:MRA是诊断AVM的有效方法,但常需结合DSA。     

Onychomycosis in Cali,Colombia

This study presents the epidemiological and mycological aspects of 299 patients with nail lesions who were referred to three diagnostic laboratories in the city of Cali. The diagnosis of mycoses was established through visualization of mycotic structures in a direct microscopic examination of skin scrapings and by isolation. Onychomycosis was found in 183 cases (61.2%), of which 141 were in toenails (44 in males and 97 in females), 38 in fingernails (9 males and 29 females), and 4 cases in toenails and fingernails simultaneously (all females). No statistically significant relation was found between sex and onychomycosis. Yeasts accounted for 40.7% of the mycoses, dermatophytes for 38%, nondermatophyte molds for 14% and the etiology was mixed in the remaining cases (7.3%). Candida albicans was the most commonly isolated yeast species; the most common dermatophyte was Trichophyton rubrum and Fusarium spp. and Scytalidium dimidiatum were the most common nondermatophytic molds. Them common fungi found in fingernails were yeasts; in toenails dermatophytes were more prevalent (chi2 with Yates' correction = 19.75, P= 0.000088). Yeasts were observed more frequently in females while dermatophytes were more common in males. The difference between these two etiologic groups was statistically significant (chi2 with Yates' correction = 7.43, P = 0.0064); no relation was observed according to age.     

Onychomycosis in Rome,Italy

This report presents the results of a study conducted between 1985 and 1994 on onychomycosis observed in the city of Rome. Six thousand six hundred and eighty eight patients were examined during this period. Among them 1,762 (26.3%) were affected by fungal nail infections. Because the etiologic agents could not be isolated in 105 cases (6%), the results refer to 1,657 subjects (24.8% of the total), presenting with positive microscopic and cultural examinations. Thirty eight patients (2.3%) had onychomycosis of both their hands and feet.From an etiological point of view, 59.1% of the nail infections were caused by yeasts, 23.2% were infected with dermatophytes and 17.6% by non-dermatophytic fungi. The etiology of onychomycosis of the hands differed from that of the feet. Yeasts were primarily responsible for onychomycosis of the hands (86.2%), while dermatophytes caused tinea unguium peduum (48%). Fungal fingernail infections by Candida spp. were the most common (50.3%), followed by those of the feet by dermatophytes (20%). Candida albicans was responsible for 70.6% of the hand infections but for only 15.9% of those of the feet. Trichophyton rubrum and T mentagrophytes were the most common dermatophytes, mainly causing toenail infections (23.4% and 21%, respectively), while Aspergillus spp., Scopulariopsis brevicaulis, Acremonium spp. and Aspergillus niger were the most common non-dermatophytes observed.With regard to sex, the fungal nail infections were more widespread in women (72.1%) and in subjects of both sexes over the age of 50.