Laboratorial aspects of cytolytic vaginosis and vulvovaginal candidiasis as a key for accurate diagnosis: a pilot study / Aspectos laboratoriais da vaginose citolítica e candidíase vulvovaginal como uma chave para o diagnóstico preciso: Um estudo piloto

Abstract Objective To identify clinical, microscopic, and biochemical characteristics that differentiate cytolytic vaginosis (CV) from vulvovaginal candidiasis (VVC). Methods The present cross-sectional study analyzed the vaginal contents of 24 non-pregnant women aged 18 to 42 years who were attended at the Genital Infections Clinic at Centro de Atenção Integral à Saúde da Mulher da Universidade Estadual de Campinas (CAISM-UNICAMP). They were diagnosed either with (CV = 8, VVC = 8) or without vulvovaginitis or vaginal dysbiosis (controls). The socio-demographic, clinical, and gynecological data were obtained from a detailed patient interview. Samples of the vaginal contents were collected for analysis of vaginal pH, gram stain, and specific fungal culture. The Kruskal-Wallis and Fisher exact tests were used to compare the differences between the groups. Odds ratios were used to compare the categorical variables. The significance level was considered at p < 0.05. Results Both women with CV and VVC had a lumpy vaginal discharge (p = 0,002) and vaginal hyperemia (p = 0.001), compared with controls. The inflammatory process was more intense in the VVC group (p = 0.001). In the CV group, there was statistical significance for the lactobacillus amount (p = 0.006), vaginal epithelium lysis (p = 0.001), and vaginal pH (p = 0.0002). Conclusion Cytolytic vaginosis and VVC diagnoses rarely differ on clinical characteristics but have different laboratorial findings. The present study highlights the importance of conducting an accurate investigation through laboratory tests rather than clinical criteria to avoid misdiagnosis.

Resumo Objetivo Identificar características clínicas, microscópicas e bioquímicas que diferenciam a vaginose citolítica (VC) da candidíase vulvovaginal (CVV). Métodos O presente estudo de corte transversal analisou o conteúdo vaginal de 24 mulheres não grávidas, com idades entre 18 e 42 anos, atendidas no ambulatório de Infecções Genitais do Centro de Atenção Integral à Saúde da Mulher da Universidade Estadual de Campinas (CAISM-UNICAMP). Elas foram diagnosticadas com (CV = 8, CVV = 8) ou sem vulvovaginite ou disbiose vaginal (controles = 8). Os dados sociodemográficos, clínicos e ginecológicos foram obtidos em uma entrevista detalhada do paciente. Amostras do conteúdo vaginal foram coletadas para análise do pH vaginal, coloração de Gram e cultura específica de fungos. Os testes exatos de Kruskal-Wallis e Fisher foram utilizados para comparar as diferenças entre os grupos. A razão de chances foi utilizada para comparar as variáveis categóricas. O nível de significância considerado foi de p < 0,05. Resultados As mulheres com VC e CVV apresentaram corrimento vaginal irregular (p = 0,002) e hiperemia vaginal (p = 0,001), em comparação aos controles. O processo inflamatório foi mais intenso no grupo CVV (p = 0,001). No grupo VC, houve significância estatística para a quantidade de lactobacilos (p = 0,006), lise do epitélio vaginal (p = 0,001) e pH vaginal (p = 0,0002). Conclusão Os diagnósticos de VC e CVV raramente diferem nas características clínicas, mas apresentam achados laboratoriais diferentes. O presente estudo destaca a importância de conduzir uma investigação precisa por meio de testes laboratoriais, em vez de critérios apenas clínicos, a fim de evitar erros de diagnóstico.

Evaluación de la coloración de Albert para la identificación de Corynebacterium spp. en flujo vaginal / Evaluation of Albert´s staining technique for Corynebacterium spp. in vaginal swabs

Introducción: la detección del género Corynebacterium en flujo vaginal representa un reto para el estudio de su posible papel patogénico en el tracto genital femenino. La coloración de Gram es la técnica recomendada para la detección de corinebacterias en muestras clínicas; no obstante, algunas características biológicas de estas bacterias dificultan la identificación por medio de esta. Objetivo: evaluar el desempeño de la coloración de Albert para la identificación de Corynebacterium spp. en muestras de flujo vaginal. Materiales y métodos: se evaluó la tinción de Albert como prueba diagnóstica para la identificación de Corynebacterium spp. en 451 flujos vaginales en comparación de la coloración de Gram. Se calculó la sensibilidad, la especificidad, los valores predictivos, las razones de verosimilitud y el índice J de Youden con el software Epidat 3.1. Resultados: la coloración de Albert presentó sensibilidad y especificidad de 61,5% y 74,6%, respectivamente, valor predictivo positivo de 78,3%, valor predictivo negativo de 56,5%, razón de verosimilitud positiva de 2,4 y negativa de 0,5 y un Índice de Youden de 0,4. Conclusiones: la tinción de Albert no mostró un mejor rendimiento que la coloración de Gram en la identificación del género Corynebacterium en muestras de flujo vaginal. Por lo tanto, se recomienda el uso en conjunto de las coloraciones de Gram y Albert para la identificación de corinebacterias en flujo vaginal. (AU)

Introduction: The detection of Corynebacterium genus in vaginal discharge represents a challenge for the study of its possible pathogenic role in the female genital tract. Gram's stain is recommended for corynebacteria detection in clinical samples, however, some biological characteristics of these bacteria difficult their identification by this technique. Objetive: to evaluate Albert's stain performans to identify Corynebacterium spp. in vaginal discharge. Materials and methods: Albert's stain was evaluated as a diagnostic test for the identification of Corynebacterium spp. in 451 vaginal swabs in comparison to Gram's stain. Sensitivity, specificity, predictive values, likelihood ratio, as well as Youden's J index were calculated using Epidat software 3.1 version. Results: Albert's stain had a sensitivity and specificity of 61.5% and 74.6%, respectively, positive predictive value of 78.3 %, negative predictive value of 56.5 %, positive likelihood ratio of 2.4 and negative likelihood ratio of 0.5, as well as Youden index of 0.4. Conclusions: Albert's stain did not show better performance than Gram's stain for identification of corynebacteria in vaginal swabs. Thereforeit is recommended the concomitant use of the Gram and Albert stains for the identification of corynebacteria in vaginal discharge. (AU)

Diagnostic value of sputum smear and simplified clinical pulmonary infection score of ventilator associated pneumonia in the early stage / 中华急诊医学杂志

Objective To explore the diagnostic value of sputum smear coupled with simplified clinical pulmonary infection score (CPIS) of ventilator associated pneumonia (VAP) in the early stage.Methods A cohort of 59 consecutive patients with VAP admitted in Intensive Care Unit from June,2014 to June,2016 were enrolled for a prospective and observational study.Concurrently,another 59 patients without pulmonary infection undergone mechanical ventilation over 48 hrs,were assigned into the control group.The criteria of exclusion were patients with pulmonary malignancies,autoimmune diseases and immunodeficiency.APACHE Ⅱ scores of all patients were recorded.All patients' inferior airway sputum which met the criteria was taken to make a validated sputum smear (i.e.polymorphonuclear leukocyte ＞ 25and squamous epithelial cell ＜ 10 per low-power field) for Gram stain and culture on the admission day.Meanwhile,simplified CPIS were calculated.Data were statistically processed by SPSS 15.0,enumeration data were statistically analyzed by Chi-Square test,and measurement data were represented as Mean ± SD.The significant differences in characteristics between two groups were analyzed by independent t test,and P ＜ 0.05 was considered statistically significant.As positive sputum smear and simplified CPIS ≥ 5 were set respectively as a positive screening criterion,sensitivity,specificity,positive predictive value and negative predictive value of each marker and combined markers were calculated.Results There were no significant differences in demographics and clinical features (including age,sex,APACHE Ⅱ scores) of patients in VAP and non-VAP patients (P ＞ 0.05).The rates of bacteria detected were Gram-negative [44.1％ (26/59)],Gram-positive [40.6％ (24/59)],none [10.2％ (6/59)] and both [5.1％ (3/59)] bacteria in VAP group,while [39.0％ (23/59)],[30.5％ (18/59)],[27.1％ (16/59)] and [3.4％ (2/59)]were found in non-VAP group correspondingly.There were no significant differences in the percentages of different bacteria in sputum smear between two groups (P ＞ 0.05).The values of diagnostic sensitivity of sputum smear and sputum smear coupled with simplified CPIS were 89.8％ and 84.7％;the specificity were 27.1％ and 79.7％;the positive predictive values were 55.2％ and 80.6％;and the negative predictive values were 72.7％ and 83.9％,respectively.Conclusions No matter the ventilated patients suffered VAP or not,bacteria might be detected from their lower respiratory tracts.Sputum smear could not be taken as an exclusively diagnostic evidence.While sputum smear coupled with simplified CPIS might improve the diagnostic efficacy of VAP,and provide the guildlines of appropriate choice of antibiotics employed in the early stage.

Comparison of five methods for the detection of fungal infections in vaginal secretions / 国际检验医学杂志

Objective To compare the similarities and differences of the five detection methods used in the detection of fungi in vaginal secretions,and find the most sensitive、the most specific、the fastest、the most cost effective and the simplest method used in the detection of fungi in vaginal secretions.Methods A total of 442 patients were selected from the Department of Gynecology of Shenzhen OCT Hospital from May 2016 to August 2016.The vaginal secretion of 442 specimens was detected by using the methods of fungi culture、saline and KOH suspension method,Gram stain,Wright''s stain and Vaginitis Multi Test Kit.In these five methods,Fungi culture were using as gold standard to evaluate the specificity,sensitivity,negative predictive value,positive predictive value and accuracy of the other four methods.Results Using the fungus culture method to detect 442 cases of vaginal secretion,we found the positive rate of mycotic infection was 34.8%(154/442).Compared with the fungi culture method,the Specificity of saline and KOH suspension method was 97.9%,the sensitivity was 64.9%,the negative predictive value was 83.9%,the positive predictive value was 94.3% and the accuracy was 86.4%;the Specificity of Gram stain was 96.5%,the Sensitivity was 83.1%,the negative predictive value was 91.4%,the positive predictive value was 92.7% and the accuracy was 91.8%;the Specificity of Vaginitis Multi Test Kit was 84.7%,the Sensitivity was 46.8%,the negative predictive value was 74.8%,the positive predictive value was 62.0% and the accuracy was 71.5%;the Specificity of Wright''s stain was 96.9%,the Sensitivity was 78.6%,the negative predictive value was 89.4%,the positive predictive value was 93.1% and the accuracy was 90.5%.Conclusion Gram stain could be the most sensitive and specific method in the four methods,with highest accuracy,and the the fastest,the most cost effective and the simplest method for the detection of fungi in vaginal secretions.The accuracy of detecting fungi in vaginal secretions could be improved by the combination of Gram stain method in clinical work.

Performance Evaluation of an Automated Microbiological Stainer, KS-S100

BACKGROUND: There have been few evaluations of the performance of automatic stainers used instead of manual staining in clinical laboratories. We evaluated the performance of an automated stainer, KS-S100 (KS Co. Ltd., Korea), in Gram and acid-fast fluorescence staining of clinical specimens. METHODS: Repeatability was evaluated by comparing the results of Gram stains performed 10 times over 5 days using two standard strains and duplicated acid-fast bacilli (AFB) fluorescence stains using 40 samples. Comparisons to manual staining and to another automatic stainer, AT-2000 (Dagatron, Korea), were performed using 40 remnant respiratory specimens or liquid AFB culture media. The results were interpreted by two experienced technicians, and total test time was measured during the staining of 1, 10, and 20 slides. RESULTS: All 10 pairs of Gram staining results obtained with KS-S100 were identical, and the concordance rate of the duplicate AFB fluorescence stains was 92.5%. Among the results for 20 culture-positive liquid media samples, three AFB fluorescent staining results were not concordant. The weighted kappa values between KS-S100 and manual staining for Gram and AFB fluorescence staining were both 0.94, with concordance within the ±1-grade range. For the staining of a single slide, manual staining was faster. KS-S100 was more efficient than AT-2000 at staining 20 slides. CONCLUSIONS: The automated stainer KS-S100 showed reproducible results comparable to those of manual staining in two major microbiological staining methods, Gram and AFB fluorescence staining, used in clinical applications. Performance evaluations should be conducted before implementation of the automated stainer in microbiological staining.

Vaginosis citolítica: presentación de un caso clínico / Cytolytic vaginosis: presentation of a clinical case

La vaginosis citolítica, descrita hace 30 años como citólisis de Döderlein, es frecuente en mujeres en la edad reproductiva y, por las características de flujo vaginal blanquecino y síntomas clínicos, es indistinguible de la vulvovaginitis micótica. Se presenta el caso de una paciente, con diagnóstico clínico presuntivo de vulvovaginitis por Candida spp. a repetición, tratada empíricamente con antifúngicos por año y medio sin ninguna mejoría. Luego de estudios microbiológicos, la coloración de Gram, demostró la presencia de 50 bacilos grampositivos por campo y abundantes núcleos celulares desnudos. El cultivo resultó puro para Lactobacillus spp., lo que permitió confirmar el diagnóstico de vaginosis citolítica. La paciente fue tratada con ampicilina-sulbactam y no ha vuelto a presentar recidivas. En conclusión, es fundamental determinar el pH vaginal de las pacientes en la consulta, así como practicar una coloración de Gram de la secreción vaginal para poner en evidencia los cambios celulares por el exceso de ácido en la vagina y así evitar tratamientos antifúngicos innecesarios que acrecentarán los trastornos de la microbiota vaginal.

Cytolytic vaginosis, described 30 years ago as Döderlein cytolysis, is common in women of reproductive age and, due to the characteristics of whitish vaginal discharge and clinical symptoms, is indistinguishable from mycotic vulvovaginitis. We describe the case of a patient with presumptive clinical diagnosis of recurrent vulvovaginitis by Candida spp. treated empirically with antifungal agents for one and a half years without improvement. After microbiological studies, Gram staining demonstrated the presence of 50 Gram-positive bacilli per field and abundant nude cell nuclei. The culture recovered pure Lactobacillus spp. which permitted the diagnosis of cytolytic vaginosis. The patient was treated with ampicillin-sulbactam and since, has not had recurrences. In conclusion, it is essential to examine the pH of the patient vaginal discharge, as well as to practice a Gram staining of the vaginal secretion to demonstrate the cellular changes produced by the excess of acid in the vagina and therefore avoid unnecessary antifungal treatments that will produce undue changes of the vaginal microbiota.

Comparison Of Objective Structured Practical Examination To Conventional Practical Examination In Microbiology.

Background & Objectives: Objective structured practical examination (OSPE) is a good tool to assess skill competency. The objectives of the study was to compare the score of marks obtained by Conventional method of assessment (CPE) with OSPE for assessing skill competency to perform Gram and Zeihl- Neelsen stain and to know the students’ and teachers’ perception about OSPE. Methods: Ninety two MBBS students were included in the study as ‘OSPE’ and ‘CPE’ groups (46+46). The two groups were assessed for their skill competency to perform Gram and Zeihl- Neelsen stain using OSPE and CPE methods respectively. Teachers’ perception was analysed for both the assessment methods with respect to objectivity, reliability, validity, feasibility etc. Feedback was taken from students too. Results: The difference of marks score by OSPE and CPE was found to be extremely statistically significant. The perception (students’ & teachers’ ) assessment reflected the acceptability of the method among students and teachers. Interpretation & Conclusion: OSPE is a tool which would help increase the objectivity while assessing skills in Microbiology and needs to be evaluated further so as to enable teachers to use this valuable tool alone or in combination with CPE.[Namrata K NJIRM 2016; 7(5):87-91]

Prevalencia de vaginosis bacteriana y factores asociados en veinte ciudades del Perú / Prevalence of bacterial vaginosis and associated factors in twenty peruvian cities

RESUMEN Objetivos. Determinar la prevalencia de vaginosis bacteriana (VB) y factores asociados en mujeres peruanas de 18 a 29 años de edad en 20 ciudades a partir de datos del proyecto PREVEN. Materiales y métodos. Estudio de tipo transversal, la definición de VB se realizó previa selección de una muestra de secreción vaginal en una lámina portaobjetos. Las láminas fueron teñidas usando la tinción Gram para ser observadas al microscopio usando el puntaje de Nugent, el diagnóstico de VB se aplicó a los puntajes 7-10. Se estimaron razones de prevalencias (RP) y sus intervalos de confianza al 95% (IC 95%) mediante el uso de modelos lineales generalizados. Resultados. Un total de 6322 mujeres contestaron la encuesta epidemiológica y proporcionaron muestras vaginales. La prevalencia de VB fue de 23,7% (IC95%: 22,6-24,7) y se asoció con tener un mayor número de parejas sexuales en los últimos 12 meses (RP: 1,22, IC 95%: 1,03-1,44, p=0,020; para dos parejas y RP: 1,46, IC 95%: 1,23-1,74, p<0,001 para tres o más parejas), no usar condón en la última relación sexual (RP: 1,16, IC 95%: 1,01-1,34, p=0,034), ser residente de la sierra (RP: 1,18, IC 95%: 1,05-1,31, p=0,004) y tener flujo vaginal anormal o con mal olor (RP: 1,20, IC 95%: 1,09-1,33, p<0,001). Conclusiones. La alta prevalencia de VB encontrada remarca la necesidad de fortalecer los servicios de salud para la detección y tratamiento de esta condición.

ABSTRACT Objetives. To determine the prevalence of bacterial vaginosis (BV) and associated factors among 18-29-year-old women in 20 Peruvian cities using PREVEN project data. Materials and Methods. In this cross-sectional study, BV was defined using previously provided vaginal discharge samples on slides, which were Gram stained and observed under a microscope to determine the Nugent scores. A BV diagnosis was applied to samples with scores of 7-10. Prevalence ratios (PR) and 95% confidence intervals (95% CI) were estimated using generalized linear models. Results. A total of 6,322 women participated in the epidemiological survey and provided vaginal swabs. The prevalence of BV was 23.7% (95% CI: 22.6-24.7) and was associated with a greater number of sexual partners in the last 12 months (PR: 1.22, 95% CI: 1.03-1.44, p=0.020 for two partners; PR: 1.46, 95% CI: 1.23-1.74, p<0.001 for three or more partners), not using a condom during last intercourse (PR: 1.16, 95% CI: 1.01-1.34, p=0.034), being a sierra resident (PR: 1.18, 95% CI: 1.05-1.31, p=0.004), and having abnormal vaginal discharge or a bad smell (PR: 1.20, 95% CI: 1.09-1.33, p<0.001). Conclusions. The high prevalence of BV highlights the need to strengthen health services aimed at the detection and treatment of this condition.

A Case of Pediatric Pertussis Unhesitatingly Diagnosed with the Insight of Microbiology Laboratory.

We describe a pediatric pertussis case occurred in an unvaccinated six-month-old male infant. The newborn patient was rapidly diagnosed by means of the insight of the clinical microbiology laboratory, despite the absence of comments on the suspicion of pertussis. The keen insight of the laboratory lay in noticing the discrepancies between the usual culture results and the gram-stain findings on the respiratory secretion submitted for bacteriological examination with the clinical diagnosis of aspiration pneumonitis. It should be highly stressed that the careful consideration of findings from gram-stained preparations surely guide correct diagnosis.

Changes in prescription of antibiotics based on Gram stains in an otorhinolaryngology outpatient clinic : A pilot study / 日本プライマリ・ケア連合学会誌

<b>Purpose</b> : This preliminary study was aimed to investigate potentially beneficial changes in prescription of antibiotics by use of Gram staining in an otorhinolaryngology outpatient clinic.<br><b>Methods</b> : This study was done in a single clinic. Changes in the type and amount of antibiotics prescribed, in addition to the clinical outcomes prior to and following introduction of Gram staining, were analyzed retrospectively.<br><b>Results</b> : Following introduction of Gram staining in 2005, the costs for antibiotics per person per year decreased to about one-fifth. The number of prescriptions per 100 patients decreased from 20.9 in 2006 to 3.6 in 2012 for macrolides, from 7.9 in 2005 to 2.4 in 2012 for third-generation cephalosporins, and increased from 1.6 in 2004 to 3.9 in 2012 for penicillins. In addition, for every 50 pediatric cases of acute sinusitis, the number of patients to whom no antibiotics were prescribed increased 9-fold and the number of patients to whom 2 or more types of antibiotics were prescribed decreased from 26 to 9 cases, while the mean period in days until resolution was shortened by 6 days.<br><b>Conclusion</b> : The results of this pilot study suggest that the use of Gram staining for diagnosis and prescription of antibiotics in an otorhinolaryngology outpatient clinic may lead to significantly improved antibiotic prescription practices. Further study through proper controlled studies are felt to be neceesary.

Interpretation of Papanicolaou Smear Test and Gram Stain Results for the Diagnosis of Infectious Vaginitis is Affected by Knowledge of Additional Related Test Results

BACKGROUND: Infectious vaginitis is a common gynecologic disease that is primarily caused by three pathogens (Trichomonas vaginalis, Gardnerella vaginalis, and Candida species). The aim of this study was to confirm the effects of other infectious vaginitis-related test results on the interpretation of Gram stain and Papanicolaou (Pap) smear test results for disease diagnosis. METHODS: A total of 300 vaginal samples were collected from women presenting symptoms of vaginitis. The presence of the three previously mentioned pathogens was evaluated using both a Gram stain and Pap smear test, and interpreted twice by 4 different observers. The first interpretation was performed without any information, and a second interpretation was performed with knowledge of results of an Affirm VPIII test that was used to diagnose infectious vaginitis. The results from the two interpretations were compared and the sensitivity and specificity of both tests were evaluated. RESULTS: For the Gram stain samples, the detection rates of G. vaginalis were increased in the second interpretation by 6.2%, while the detection rates of Candida spp. were decreased by 0.3%. For the Pap smear test samples, the detection rates of G. vaginalis were increased in the second interpretation by 7.0%, and the detection rates of Candida spp. were increased by 2.0%. The sensitivity of both tests was increased in the second interpretation by 5.5% to 66.7%. There was no difference in the specificity between the two interpretations. CONCLUSIONS: We demonstrated that there is significant inter-observer variation when using Gram stain and Pap smear test results to diagnose infectious vaginitis. The detection rates and sensitivity of both tests changed when the results from an additional test were incorporated into the interpretation. Additional studies are needed to develop objective criteria and a standardized interpretation system for the evaluation of results from these diagnostic tests.

Pap Smears in the Diagnosis of Bacterial Vaginosis.

Pap smears are routinely used in cervical cancer screening. The purpose of our study was to determine whether the Pap smear is of diagnostic value for the detection of bacterial vaginosis using vaginal gram stain as the diagnostic standard. We studied 636 non-pregnant, married women attending the Gynecology OPD of Rama Medical College, Ghaziabad, for routine genital examination between Sep 2011 and Sep 2012. Two vaginal smears were prepared, one of which was stained by the pap method and the second by the Gram’s method. Using Gram stain diagnosis of bacterial vaginosis as the gold standard, pap smear had a sensitivity of 61.0% and a specificity of 97.6%, PPV of 85.7% and NPV of 91.6%in the diagnosis of bacterial vaginosis. Hence, although pap smear is only moderately sensitive for screening of bacterial vaginosis, because of its high specificity, it is of diagnostic value when it is positive.

Tinción de Gram de tejido: alcances y limitaciones / Gram stain of tissue biopsy: scope and pitfalls

La identificación de microorganismos en tejido es esencial para reconocer un proceso infeccioso. Inicialmente, mediante la coloración de hematoxilina-eosina, se puede identificar el patrón de inflamación asociado y luego, a través de tinciones basadas en plata y la tinción de Gramde tejido se visualizan los microorganismos. La tinción de Gram no solo sirve para bacterias, sino también para algunos hongos y parásitos; no obstante, esta técnica tiene algunos inconvenientes, como la contaminación con otros microorganismos y la imposibilidad de visualizar algunas bacterias, entre ellas Legionella pneumophila, Leptospira spp y Bartonella spp. En este artículode revisión se describirán los fundamentos del Gram de tejido, su contribución en el diagnóstico de infecciones como herramienta adicional para el reconocimiento de microorganismos, y sus limitaciones.

The identification of microorganisms in tissue is pivotal to recognize infectious processes.At first, the hematoxylin-eosin stain is used to identify the pattern of inflammation associated; after that, microorganisms are seen through Gram or silver stains. Gram Stain of tissue biopsy not only stains bacteria, but also a number of fungus and parasites. However, this technique has some disadvantages, such as contamination with other microorganisms, and lack of stain of some bacteria, including Legionella pneumophila, Leptospira spp and Bartonella spp. This review articleaims to describe the fundamentals of Gram stain of tissue biopsy and its assistance in infectious diagnosis as an additional tool for recognition of microorganisms, as well as its limitations.

Detection of Trichomonas vaginalis, Gardnerella vaginalis, and Candida Species in Affirm VPIII, Papanicolaou Smear Test and Gram Stain / 대한임상미생물학회지

BACKGROUND: Infectious vaginitis is caused primarily by three different groups of microbial pathogens (Trichomonas vaginalis, Candida spp., and Gardnerella vaginalis). The objective of this study was to compare the Affirm VPIII assay using a DNA hybridization technique with the Papanicolaou (Pap) smear test and the Gram stain in the detection and identification of these three organisms. METHODS: A total of 300 vaginal samples were collected from women that were either symptomatic for vaginitis or asymptomatic women that were being seen for routine obstetric or gynecological care. The presence of T. vaginalis, Candida spp., and G. vaginalis was evaluated by using the Affirm VIII assay (Becton Dickinson, USA), Pap smear test, and Gram stain method, respectively. RESULTS: With the Affirm VPIII assay, 1 (0.3%) patient tested positive for T. vaginalis, 99 (33.0%) patients were positive for G. vaginalis, and 18 (6.0%) were positive for Candida spp. The detection rates of Trichomonas infection, bacterial vaginosis and candidiasis by the Pap smear test and Gram stain method were 0.7% versus 0%, 16.3% versus 35.7%, and 1.7% versus 9.7%, respectively. The differences between the detection rates of the above three organisms between the Pap smear test and the Gram stain method were statistically significant (p<0.05). CONCLUSION: The Affirm VPIII assay was more sensitive than the Pap smear test and more specific than the Gram stain method for the detection and identification of these three organisms. In addition, the results of the Affirm VPIII assay are quick to obtain and are simple and easy to interpret.

Significance of direct smear examination for the diagnosis of female genital tract infections.

The present study is conducted to demonstrate the value of direct smear examinations ( Gram stain and wet mount ) of vaginal discharge from infected female patients. The diagnosis of bacterial vaginosis and other infections were carried out by direct gram stained smear and wet mount preparations of vaginal discharge in 1004 cases of various infections affecting these patients. Gram stain was a useful tool to pick up various infections of the genital tract and wet mount was helpful in detecting Trichomonas vaginalis. In short , direct smear examination will facilitate clinician in the diagnosis of female genital tract infections with minimum requirements.

Comparación de los criterios de Nugent y Spiegel para el diagnóstico de vaginosis bacteriana y análisis de los resultados discordantes por el método de Ison y Hay / Nugent and Spiegel criteria for the diagnosis of bacterial vaginosis. Analysis of discordant specimens by the Ison and Hay method

Background: Vaginal infection is the commonest cause of genital symptoms and has obstetric and gynecological implications. Aim: To compare the Nugent and Spiegel methods for the diagnosis of bacterial vaginosis (BV) and to analyze discordant specimens using Ison and Hay (Ison/Hay) criteria. Material and Methods: After discardingcases with Candidiasis, deficientspecimens or those lacking bacteria, a total of348 Gram-stained smears vaginal specimens receivedfor the diagnosis of BV, were analyzed. Results: Vaginal microbiota was classified as normal in 203 and 237 samples (58 and 68 percent of samples), accordingto Nugent and Spiegel criteria, respectively One hundred andfive (30 percent) and 111 samples (32 percent), were classified as VB accordingto Nugent and Spiegel criteria, respectively. Both criteria were concordant in 308 samples (88.5 percent). The 40 (11.5 percent) discordant specimens were classified as intermedíate microflora by the Nugent system and as normal or BV by Spiegel. Among these, the Ison/Hay procedure identified four categories of microbiota. Ten (25 percent) specimens were classified as grade II microbiota, confirming their categorization by Nugent as intermedíate microbiota, six (15 percent) were classified in the III category, confirming the diagnosis ofBV by Spiegel, 13 (32.5 percent) corresponded to the category III, that does not exist in the Nugent and Spiegel categorization systems. Finally, 11 specimens could not be assigned to one category due to microscopic limitations to distinguish bacterial morphotypes. Conclusions: The systems proposed by Spiegel, Nugent and Ison I Hay are comparable for the diagnosis of BV. However, we recommend the use of Ison/ Hay procedure to evalúate vaginal microbiota, due to its wider range of categories, allowing a better discrimination ofthe vaginal microbiota.

Blood culture gram stain, acridine orange stain and direct sensitivity-based antimicrobial therapy of bloodstream infection in patients with trauma.

Purpose: The purpose of this study was to ascertain if the simple practice of Gram stain, acridine orange stain and direct sensitivity determination of positive blood culture bottles could be used to guide early and appropriate treatment in trauma patients with clinical suspicion of sepsis. The study also aimed to evaluate the error in interpreting antimicrobial sensitivity by direct method when compared to standard method and find out if specific antibiotic-organism combination had more discrepancies. Findings from consecutive episodes of blood stream infection at an Apex Trauma centre over a 12-month period are summarized. Materials and Methods: A total of 509 consecutive positive blood cultures were subjected to Gram staining. AO staining was done in BacT/ALERT-positive Gram-stain negative blood cultures. Direct sensitivity was performed from 369 blood culture broths, showing single type of growth in Gram and acridine orange staining. Results of direct sensitivity were compared to conventional sensitivity for errors. Results: No 'very major' discrepancy was found in this study. About 5.2 and 1.8% minor error rates were noted in gram-positive and gram-negative bacteria, respectively, while comparing the two methods. Most of the discrepancies in gram-negative bacteria were noted in β lactam - β lactamase inhibitor combinations. Direct sensitivity testing was not reliable for reporting of methicillin and vancomycin resistance in Staphylococci. Conclusions: Gram stain result together with direct sensitivity testing is required for optimizing initial antimicrobial therapy in trauma patients with clinical suspicion of sepsis. Gram staining and AO staining proved particularly helpful in the early detection of candidaemia.

Interobserver variation in the interpretation of Nugent scoring method for diagnosis of bacterial vaginosis.

Background & objectives: Bacterial vaginosis (BV) is the most common form of vaginal infection and an important cause of morbidity in women of reproductive age. This study was carried out to examine the interobserver variation on interpretation of Nugent scoring method in the diagnosis of BV. Methods: This prospective study was conducted in a rural primary health care center of north India from May 2003 to April 2004 and included 601 married, sexually active women between 18-49 yr of age presented with self-reported symptoms of vaginal discharge and/or genital itching and/or genital burning. Specimens collected from the lateral wall of vagina were subjected to Gram staining and the microscope slide smears were examined by 3 independent observers. Each of the three observers scored and interpreted the slides for diagnosis of bacterial vaginosis using the Nugent method. Results: Complete agreement amongst the three observers was found in 76.2 per cent of cases. In 22.13 per cent cases, two observers were in agreement while interpretation of the slides were in complete disagreement only in 1.66 per cent of cases. The interrater reproducibility was found to be excellent between observers 1 and 3, while between observers 1 and 2, and 2 and 3 it was good to fair. Interpretation & conclusions: Nugent scoring system appears to be a reliable and convenient method for laboratory evaluation of cases of bacterial vaginosis. At the same time, one must be aware of the factors that might lead to discrepant results.

Clostridium symbiosum Isolated from Blood / 대한임상미생물학회지

Clostridium symbiosum was isolated from the blood of a 61-year-old immunocompromised woman who had diagnosed ovarian cancer with multiple metastases and who had developed persistent tachycardia. A blood culture was drawn from the peripherally inserted central catheter, and non-spore-forming gram-negative rods were detected in an anaerobic vial. The organism showed tiny and pinpoint colonies and was unidentified by Vitek II (bioMerieux, France). The 16S rRNA gene sequence showed a 99.4% identity with C. symbiosum. To our knowledge, this represents the first report of C. symbiosum isolation in Korea.

Cambios del esquema antibiótico en neumonía adquirida en la comunidad. Valor del cultivo de esputo en pacientes ingresados al Hospital de San José, Bogotá D.C. / Changes in antimicrobial regimes to treat community-acquired pneumonia: value of sputum culture in patiens admitted to the San José Hospital, Bogotá D. C.

Introducción: la utilidad del Gram y cultivo de esputo para el manejo y seguimiento de los pacientes con neumonía adquirida en la comunidad (NAC) es aun controversial. Objetivo: describir la frecuencia de cambio del manejo antibiótico de acuerdo con el resultado del cultivo de esputo en enfermos hospitalizados con NAC. Tipo de estudio: descriptivo de corte transversal. Métodos: pacientes ingresados al servicio de medicina interna del Hospitalde San José con diagnóstico clínico y radiológico de NAC que no requirieran manejo en UCI. Se incluyeron 86 casos en un período de cuatro meses a quienes se les aplicó al ingreso la escala de pronóstico CURB 65, la presencia o no de comorbilidades, la positividad del Gram y cultivo de esputo, la estancia hospitalaria y el cambio de antibiótico de acuerdo con el resultado del cultivo de esputo si la muestra era significativa. Resultados: el promedio de edad de la población fue 58.8 años (DS 21.7). El 45% tenía comorbilidades al ingreso y el 37% algún tipo de complicación asociada con la NAC. En la escala de CURB-65 el 52% tenía un punto, dos puntos el 27%, tres el 3.5 % y con cuatro el 2%. La respuesta a la terapia antibiótica se observó en el 85% y la frecuencia del cambio antibiótico según el reporte de cultivo ocurrió en el 10% (6/62). La mediana de días de estancia hospitalaria fue de cinco. El 95% egresó del hospital y el 5% restante fue trasladado a otro servicio. Conclusiones: el Gram y cultivo de esputo son ayudas diagnósticas de bajo costo y útiles en pacientes con NAC clasificados segúnlas escalas de severidad de alto riesgo o neumonías severas, en donde el aislamiento del germen es crucial paradirigir la terapia antibiótica; en neumonías no severas no han demostrado mayor utilidad ya que pocas veces hay aislamiento del germen y no generan cambios en el manejo del paciente. Se debe asegurar la adecuada toma y procesamiento para aumentar la probabilidad de aislamiento.

Introduction: the applicability of sputum Gram stain and culture to define management and follow-up of patients with community-acquired pneumonia (CAP) remains controversial. Objective: to describe the frequency of changes in antimicrobial therapy based on results of sputum culture in hospitalized patients with CAP. Type of study: this is a descriptive cross sectional study. Methods: patients admitted to the Internal Medicine Department at the San José Hospital with radiologic and clinic diagnosis of CAP who did not require ICU care were selected. Eighty-six (86) cases in a four-month period were included. They were graded with the CURB-65 prediction scale upon admission considering comorbid factors, positive sputum Gram stain and culture, hospital stay and change of antimicrobial therapy based on the result of the sputum culture when sample was substantial. Results: the mean age of the study population was 58,8 years (SD 21,7). Forty-five percent (45%) had comorbidities upon admission and 37% had some type of complication associated to CAP. In the CURB-65 scale 52% rated one point, 27% rated two points, 3,5 % three points and 2% four points. Response toantimicrobial therapy was observed in 85% and a change of antimicrobial regime based on culture results occured in 10% (6/62). The median number of days in hospital was five. Ninety-five percent (95%) were discharged from the hospital and the remaining 5% were transferred to another department. Conclusions: sputum Gram stain and culture are low-cost diagnostic tools and are useful in patients with CAP classified acording to the high-risk severity scales or severe pneumonias, in which isolation of the pathogen is crucial to tailor antimocrobial therapy; in non-severe pneumonias they have not shown mayor applicability for in a very small number of cases the etiologic organism is isolated and no changes in patient management are generated...