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Background: Taenia solium infections in humans include the infection by the adult tapeworm, these infections are of public health concern and are among the most important afflictions of humans who live in areas of poverty in the developing world and least developed countries. T. solium, a zoonotic disease, transmitted between pigs and humans and among humans, is common in developing countries. Aims and Objectives: The aim of the study was to estimate the detection rate of T. solium taeniasis among patients and random community screening with an indication of intestinal parasitic infection by routine stool examination. Materials and Methods: Stool samples were collected from the community and patients. Those who were willing, samples were screened for the cysts/ova/egg by direct microscopic examination by saline, iodine, concentration technique, and modified acid fast staining, were performed to differentiate species of T. solium and Taenia saginata. Results: Overall samples were 2030, out of which 870 stool samples were from community field screening 585 (28.81%) were positive. 1160 from tertiary care center, 668 (32.90%) were positive gave a total prevalence of intestinal parasitic infection of 61.72%. The prevalence of T. solium taeniasis was 194 (9.55%) out of which 92 (4.53%) were from community and 102 (5.02%) were from tertiary care center. Conclusion: The high prevalence of intestinal parasitic infestation might be due to the poor sanitary, contaminated water, and lack of education that is prevalent in the studied region as in other pockets in rural India. Our study showed the usefulness of the Ziehl-Neelsen modified acid-fast stain for identification of Taenia species.
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INTRODUCCIÓN: Tuberculosis (TBC) sigue siendo la segunda causa de muerte por una enfermedad infecto-contagiosa después del síndrome de inmunodeficiencia humana adquirida (SIDA). Actualmente, el escenario de técnicas y metodologías de laboratorio para la identificación y drogo-sensibilidad está cambiando gradualmente. Se han recomendado e introducido ensayos rápidos basados en la amplificación de ácidos nucleicos (NAAT's) que se desarrollan mediante la reacción de polimerasa en cadena (RPC). Bajo este principio, se destaca spoligotyping -una herramienta de genotipificación y epidemiología molecular en TBC- estandarizada a partir de aislados bacterianos, que permite el estudio del genoma de Mycobacterium mediante la amplificación de 43 secuencias cortas no repetitivas, localizadas en la región de repetición directa (RD1). OBJETIVO: Evaluación de spoligotyping a partir de baciloscopías, como una opción independiente de cultivo, para la caracterización de Mycobacterium tuberculosis a partir de muestras de esputo en pacientes del Instituto Nacional Cardiopulmonar de Tegucigalpa, Honduras. MÉTODO: De 37 pacientes con cultivo (y baciloscopía) positivos para M. tuberculosis, se obtuvieron 50 muestras de expectoración. Se realizó estudio microbiológico y molecular en muestras respiratorias conteniendo ADN de micobacterias, a partir de baciloscopías, concentrados y cultivo, para la identificación y análisis genotípico a través de la técnica de spoligotyping. RESULTADOS: El spoligotyping fue positivo en 37/37 de muestras de cultivo positivo (S: 100%), en 36/37 (S: 97,3%) de muestras con baciloscopía positiva y en 6/10 (S: 60%) de muestras de concentrado de esputo. La intensidad de la baciloscopía positiva tuvo una relación directa con la sensibilidad de spoligotyping. DISCUSIÓN: El fusionar el potencial de una herramienta útil en epidemiología molecular para analizar muestras de ADN proveniente de baciloscopías, visualiza una plataforma diagnóstica y genotípica para países en vías de desarrollo como una alternativa innovadora y altamente sensible en la hibridación de oligonucleótidos específicos a partir material genético en baciloscopías (P+, P++, P+++), pero requiere mejorar la concordancia entre patrones genéticos obtenidos, comparables con el uso estandarizado de aislados de cepas de M. tuberculosis.
BACKGROUND: Tuberculosis (TB) remains as the second cause of death by an infectious disease preceded by the acquired immune deficiency syndrome (AIDS). Currently, laboratory techniques and methodologies of diagnosis and drug susceptibility testing are constantly changing. Therefore, it has been recommended the introduction of rapid assays based on the amplification of nucleic acids test (NAAT's) through a polymerase chain reaction (PCR). Based on this principle, outstands spoligotyping - as a genotype and molecular epidemiology tool in tuberculosis - it is standardized to use isolated bacteria for the study of Mycobacterium genome through the amplification of 43 non-repetitive sequences, located at the direct repetitive region 1 (RD1). AIM Evaluation of spoligotyping from acid fast staining smears as an independent option from bacterial isolation to characterize Mycobacterium tuberculosis by using sputum samples from TB patients from National Cardiopulmonary Institute in Tegucigalpa, Honduras. METHOD: Of 37 patients with positive culture (and smear microscopy) for M. tuberculosis, 50 expectoration samples were obtained. Microbiological and molecular tests were performed in respiratory samples containing mycobacterial DNA from sputum smears, concentrates and solid culture, for identification and genotype analysis by spoligotyping technique. RESULTS: Spoligotyping was positive in 37/37 of positive culture samples (S: 100%), in 36/37 (S: 97.3%) of smear-positive samples and in 6/10 (S: 60%) of concentrate samples sputum. The intensity of positive smear microscopy had a direct relationship with the sensitivity of spoligotyping. DISCUSSION: This study combined the potential of a molecular epidemiology tool to analyse DNA from sputum samples in smears acid fast staining, it visualizes diagnosis and genotyping platform in developing countries gathering innovation and high sensitivity in the hibridization of specific olignonucleotides from positive smears (P+, P++, P+++). However, the low specificity showed the need to improve better agreement among genetic patterns compared to the standardized bacterial isolation from M. tuberculosis strains.
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Humans , Tuberculosis/diagnosis , Mycobacterium tuberculosis/genetics , Sputum , Microbial Sensitivity Tests , Sensitivity and Specificity , GenotypeABSTRACT
Objective To explore the different diagnostic values of acid-fast staining, tuberculosis culture, tuberculosis DNA detection (TB-DNA), tuberculosis RNA constant temperature amplification technology (SAT-TB) in the detection of Mycobacterium tuberculosis in sputum specimens. Methods A total of 200 pulmonary tuberculosis patients and 80 non-tuberculosis patients who were hospitalized in Hebei Chest Hospital between September 2015 and September 2019 were selected for this study. Sputum samples were collected after admission, and the detection values of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB in sputum samples were statistically analyzed. Results The differences in the sensitivity, accuracy, positive predictive value, and negative predictive values of the four diagnostic methods of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB were statistically significant (P TB-DNA> tuberculosis culture> acid-fast staining. In terms of the positive predictive value of diagnosis, the values of SAT-TB, TB-DNA, and tuberculosis culture were higher than that of acid-fast staining. The Kappa values of the four methods and the gold standard were: Kappa (acid-fast staining) = 0.145, Kappa (tuberculosis culture) = 0.395, Kappa (TB-DNA) = 0.602, and Kappa (SAT-TB) = 0.770. Conclusion The four diagnostic methods of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB all had certain detection value with their advantages and disadvantages. SAT-TB was a better detection method with high specificity, good sensitivity, and a short detection timer, which could quickly identify bacteria and distinguish live bacteria.
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Intestinal parasitic infections is a serious public health problem in most of the regions of the world, especially in developing countries, and represents a major cause of morbidity and mortality in children and among high-risk groups. Aims: To find out the prevalence of intestinal parasitic infections in Garhwal region of Uttarakhand and compare and correlate it with gender, age group and area (rural or urban). Methods: The collected stool samples were subjected to routine stool investigations during the study, i.e. Macroscopic examination was carried out for the presence adult worms or their body segments and microscopic examination such as stool wet mounts (both saline and iodine mounts); modified acid-fast staining for Cryptosporidium and Isospora and ELISA for Cryptosporidium were done. Results: Out of total 3614 patients, 197 (5.45%) (excluding Cryptosporidium ) and 338(9.35%) (Including Cryptosporidium), while 141(3.90%)(only Cryptosporidium )had parasitic infection. Maximum numbers (average) of patients were enrolled in month of May (102, 16.94%) and June (87, 14.45%). Maximum number of patients were in the age group of 1-10yrs (949, 26.26%) whereas out of these110 patients were found positive (11.6%). On the other hand, out of 343 patients in 11-20yrs age group, 46 (13.4%) were found positive and least number of patients were from <1year age group. Mostly male patients were affected by parasitic infections (60.06%). Parasite most commonly isolated was Cryptosporidium 141(41.72%), followed by Giardia lamblia 74(21.89%) and hookworm 40(11.83%). One cases each of trematodes, Fasciola hepatica (both ova and adult) and Clonorchis (ova) whereas two cases of Isospora belli were also seen. Conclusion: Prevalence of intestinal parasites (9.35%) is low in Garhwal region of Uttarakhand.
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Objective To investigate the status of Cryptosporidium infection in the population in Nanjing City so as to pro-vide the evidence for the prevention and control of cryptosporidiosis. Methods A total of 100 fecal samples were collected from each of three districts(Liuhe,Qixia and Gaochun)and one hospital(Nanjing Zhongda Hospital)in 2015 and 2016 respective-ly. The fecal samples were detected for Cryptosporidium with microscopy(by using the gold amine phenol-modified acid-fast staining)and the positive samples were detected again for the molecular biology confirming by using the fluorescence quantita-tive PCR. Results During the two years,581 cases of normal population who lived in the city were surveyed and no Cryptospo-ridium infection was found. Among 202 cases of outpatients with chronic diarrhea,there were 9 Cryptosporidium positive cases with the microscope scanning method (4.46％),and among the 9 cases,7 cases showed obvious logarithmic amplification curves showing positive Cryptosporidium nucleic acid,but 2 cases without the obvious logarithmic amplification curves,and the Cryptosporidium nucleic acid positive rate was 3.47％. Conclusions Cryptosporidium infection is not found in the normal popu-lation of Nanjing City,but the Cryptosporidium infection is found in the chronic diarrhea patients. The results imply that we should strengthen the detection of Cryptosporidium in the chronic diarrhea patients,so as to provide the evidence for improving the diagnosis and treatment of cryptosporidiosis.
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Background and Aim: Cutaneous tuberculosis (CTB) is still difficult to diagnose due to its varied clinical presentation and limitations of diagnostic methods. The aim of this study was to evaluate the results of diagnostic laboratory tests available for CTB. Materials and Methods: Twenty-six skin biopsy specimens belonging to clinically suspected cases of CTB were studied retrospectively. The specimens were divided into two portions, one part processed for histopathological evaluation and the other was used for microscopy and inoculation for the isolation of mycobacteria. Polymerase chain reaction (PCR) technique was applied to 14 of 26 specimens to detect Mycobacterium tuberculosis complex (MTBC) DNA. Results: Of the 26 biopsy specimens, 11 were confirmed as CTB by identification of MTBC in culture and/or histopathologic affirmation. Of these, four were lupus vulgaris, four were TB verrucosa cutis, one was scrofuloderma, one was primary inoculation TB, and one was periorifical CTB. Culture for mycobacteria was positive for five (45.45%) specimens, while histopathologic affirmation was obtained in ten (90.90%) specimens. Acid-fast Bacilli were not demonstrated in any of the specimens on microscopic examination. The PCR was found to be applied to six of the 11 specimens diagnosed as CTB and was positive in two specimens (33.3%), which were positive for growth in culture and histopathological correlation. Conclusion: The recovery rate of MTBC from biopsy specimens was found to be satisfactory for CTB with histopathological correlation, but the combination of culture with a rapid method, PCR, may improve the diagnostic rate.
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Objective To evaluate the value of four methods in diagnosis of pulmonary tuberculosis,including T-SPOT,fluo-rescent PCR,anti-TB antibody test,and acid-fast staining.Methods Retrospective analysis of 530 cases between January 2012 and December 2014 who had taken four methods,and calculated the sensitivity specificity,positive predictive value,neg-ative predictive value,Kappa value,Youden index,positive likelihood ratio,negative likelihood ratio,Pairedχ2 test.Consider clinical diagnosis as the gold standard.Results The sensitivity,negative predictive value,Youden index of T-SPOT were 95.90%,97.29%,0.82,respectively,and all of these were the highest.The negative likelihood ratio of T-SPOT was 0.05, which was the lowest.Misdiagnosis rate of PCR was 87.18%,which was the highest.Positive likelihood of anti-TB antibody test was the lowest,6.48,while other indicators were no advantage.The specificity,positive predictive value and positive likelihood ratio of acid-fast staining were 99.70%,98.90%,153.83,respectively,and the three of these were highest.Pair-wise comparison between the four methods were significantly different.Conclusion The T-SPOT and acid-fast staining can be used as important methods,and the anti-TB antibody can provide result quickly,and PCR method is more suitable for ex-amination of sterile body fluids.
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Objective To compare the clinical effect of application of gene chip microscopy technique for rapid identification of Mycobacterium and classic smear acid-fast staining,and to assess the advantages and disadvantages of the wo methods.Methods From 201 1 to 2014,gene chip microarray and smear acid fast staining were used to identify the mycobacterium tuberculosis in speci-mens suspicious of the infection from all the general hospitals of Shenzhen city.Chi-square test was used to compare the positive rates of the two methods.Results A total of 2 481 specimens were collected from clinic.With smear acid-fast staining technique, the positive specimens of 1 93 cases werefound and the positive rate was 7.8%.Meanwhile,31 7 positive samples were detected by the technology of gene chip microarray,and the positive rate was 12.8%.The positive rate of Gene chip microarray technology was higher than that of the smear acid fast staining,and there was significant difference between them (P < 0.05 ).The 31 7 positive samples identified by Gene chip microarray,included 263 cases of Mycobacterium tuberculosis,27 cases of Mycobacterium absces-sus,18 cases of Mycobacterium intracellulare,3 cases of Mycobacterium gastric uLcer,3 cases of Mycobacterium avium,1 case of Mycobacterium Gordonae,1 case of Mycobacterium marinum and 1 case of Mycobacterium Kansas.Conclusion The gene chip mi-croarray technology is fast,accurate,and its positive rate is higher than that of smear acid-fast staining technique.Classification and identification of Mycobacterium is very helpful for clinical individualized treatment of anti mycobacterium infection.
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Objective To investigate the status of Cryptosporidium infection in the population in the local area of Anhui Prov?ince,and discuss the risk factors of the infection,so as to provide the evidence for the prevention and treatment of cryptosporidi?osis. Methods Qianshan County and Lingbi County of Anhui Province were selected as investigation spots,and the oocysts of Cryptosporidium in the feces of the investigation objects and the specific IgG antibody against Cryptosporidium in the serum were checked by using the pathogenic modified acid fast staining method and ELISA,respectively,so as to determine the status of Cryptosporidium infection in these investigation objects. At the same time,the questionnaire surveys were conducted in the inves?tigation objects so as to know about the risk factors of Cryptosporidium infection. Results A total of 668 people were investigat?ed in the two counties,635 people received etiological examinations,and 15 people were positive with the positive rate of 2.36%;642 people received serological examinations,and 140 people were positive with the positive rate of 21.81%;628 peo?ple received pathogenic and serological examinations at the same time,and the examination results of the both methods showed that 12 people were positive(there were 4 people in Qianshan County and 8 people in Lingbi County),and the positive rate was 1.94%. The rates of Cryptosporidium infection in the population of Qianshan County and Lingbi County were 1.24%(4/322)and 2.71%(8/295)respectively,and the difference had no statistical significance(P>0.05). The single factor analysis found that the rate of Cryptosporidium infection was higher in the children and diarrhea patients;the multivariate logistics regression analy?sis indicated that the rate of Cryptosporidium infection was higher in the people who bred poultry and the diarrhea patients. Con?clusions The positive rate of serum antibody of Cryptosporidium in the population of the local area of Anhui Province is higher, which indicates that the previous infection is serious,and the rate of Cryptosporidium infection in human is relative to the age, diarrhea and whether there are poultries to be bred in the family,which is worthy of attention in the future prevention and treat?ment.
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Objective To compare the results difference between the fluorescence staining and the acid fast staining (Ziehi‐Neelsen staining) methods in the detection of Mycobacterium tuberculosis ,and to compare the effects of the methylene blue solu‐tion ,Haris hematoxylin solution and potassium permanganate liquid as the redyeing reagents of the fluorescence staining method . Methods 198 sputum specimens collected from the patients with suspected tuberculosis symptoms and were performed the Ziehi‐Neelsen staining and the fluorescence staining respectively For comparing the difference in the detecting rate of Mycobacterium tu‐berculosis between the two kinds of method .The fluorescence staining adopted 0 .3% methylene blue solution ,0 .5% Haris hema‐toxylin solution and 0 .5% potassium permanganate solution as the redyeing reagents for comparing the effects of the fluorescence microscopic examination among different redying reagents .Results The detection rate of Mycobacterium tuberculosis was 66 .67%(132/198) for the Ziehi‐Neelsen staining ,94 .9% (188/198) for the fluorescence stainings and 94 .95% (188/198) for the methyl‐ene blue staining ,in which the detection rate of methylene blue redyeing was 94 .95% ,which of hematoxylin redyeing was 94 .44%(187/198) and which of potassium permanganate redyeing was 94 .44 (187/198) ,the differences among them were statistically sig‐nificant(P< 0 .05) .Conclusion The fluorescent staining method has the higher positive detection rate of Mycobacterium tubercu‐losis than the Ziehl‐Neelsen staining method ,in which 0 .3% methylene blue solution is a good background quenching redyeing solu‐tion .
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OBJECTIVE: To describe laboratory personnel's attitude and practices toward phenol exposure during Ziehl Neelsen (ZN) acid fast staining method and to evaluate the feasibility of an alternate modified Kinyoun cold (MKC) stain. METHOD: A total of 187 sputum samples were collected from suspected tuberculosis cases and stained by the MKC method and ZN stain and were read by an experienced microscopist and a researcher. A crosssectional questionnaire survey of 35 laboratory personnel was also conducted. RESULTS: Modified Kinyoun cold stain gave sensitivity, specificity, positive and negative predictive values of 100%, 99.4%, 94.1% and 100%, respectively. Both stains corresponded with an agreement rate of 99.5%. Almost 94.7%of respondents reported that they worked in a closed area when staining and 57.1% did the staining method without ventilation. Material safety data sheet (MSDS) of phenol was not known to 77.1% of laboratory personnel. All of the participants (100%) in this study welcomed a similar, non heating method for acid-fast bacillus (AFB). There was significant association between those not comfortable with phenol exposure (77.1%) and complaints of irritation (48.6%) and headache (2.9%) [χ² = 10.98, r = 0.55, p = 0.001]. CONCLUSIONS: The MKC is suitable for use as a substitute for the ZN method for the demonstration of AFB in the primary diagnosis and treatment assessment of pulmonary tuberculosis. Focus should be given on educating laboratory staff on the hazards, risks and precautions associated with the phenol/ZN method.
OBJETIVO: Describir las actitudes y prácticas del personal del laboratorio hacia la exposición al fenol durante la aplicación del método de tinción ácido-rápida de Ziehl Neelsen (ZN), y la viabilidad de la alternativa de una tinción de Kinyoun en frío modificada (MKC). MÉTODO: Un total de 187 muestras de esputo fueron recogidas de casos con sospecha de tuberculosis, y teñidas por el método MKC y la tinción de ZN, tras lo cual fueron leídas por un microscopista y un investigador con experiencia. También se realizó una encuesta transversal a manera de cuestionario, entre las 35 personas que conformaban el personal del laboratorio. RESULTADOS: La tinción de Kinyoun en frío modificada arrojó sensibilidad, especificidad, y valores predictivos positivos y negativos de 100%, 99.4%, 94.1% y 100%, respectivamente. Ambas tinciones se correspondieron con una tasa de acuerdo de 99.5%. Casi el 94.7% de los encuestados informó que trabajaban en un área cerrada en el momento de la tinción, y 57.1% tuvo el método de la tinción sin ventilación. La ficha de datos de seguridad (FDS) de fenol era desconocida para el 77.1% del personal de laboratorio. Todos los participantes (100%) en este estudio dieron la bienvenida a un método similar, sin calentamiento, para los bacilos acidoresistentes (BAR). Hubo una asociación significativa entre aquellos para quienes la exposición al fenol les era inconfortable (77.1%) y las quejas de irritación (48.6%) y dolor de cabeza (2.9%) [χ² = 10.98, r = 0.55, p = 0.001]. CONCLUSIONES: El MKC es adecuado para uso como sustituto del método de ZN para la demostración de BAR en el diagnóstico primario y la evaluación del tratamiento de la tuberculosis pulmonar. Debe centrarse la atención en educar al personal de laboratorio en los peligros, los riesgos, y la precaución asociados con el método de fenol/ZN.
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Humans , Staining and Labeling/methods , Health Knowledge, Attitudes, Practice , Occupational Exposure/prevention & control , Medical Laboratory Personnel , Phenol/adverse effects , Coloring Agents/adverse effects , Sputum/microbiology , Tuberculosis, Pulmonary/diagnostic imaging , Cross-Sectional Studies , Surveys and Questionnaires , Sensitivity and SpecificityABSTRACT
Fecal samples of 2,056 dairy cattle from 14 farms were collected in three geographical regions of China and stained using a modified acid-fast staining technique to identify Cryptosporidium oocysts. A total of 387 (18.82%) positive samples were identified and further analyzed by polymerase chain reaction (PCR) using primers designed to amplify DNA fragments from the small subunit ribosomal RNA. The PCR products were sequenced and the sequences were deposited in the GenBank database under accession numbers EU369377-84 and GU070730-33. Phylogenetic analysis was performed and a distances matrix generated from these sequences confirmed the existence of Cryptosporidium (C.) parvum 'mouse' genotype, C. bovis, C. andersoni, C. hominis, and C. serpentis in cattle. These results represent the first report on the prevalence and genetic identification of Cryptosporidium species, and may contribute to a better understanding of the epidemiology of Cryptosporidium in cattle in China.
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Animals , Cattle , Female , Base Sequence , Cattle Diseases/epidemiology , Chi-Square Distribution , China/epidemiology , Cryptosporidiosis/epidemiology , Cryptosporidium/genetics , DNA, Protozoan/chemistry , Feces/parasitology , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction/veterinary , Prevalence , RNA, Ribosomal, 18S/chemistry , Sequence Alignment , Sequence Analysis, DNAABSTRACT
Objective To explore the relationship between chronic severe hepatitis and cryptosporidium infection to provide evidences for scientific prevention and treatment of chronic severe hepatitis. Methods Fecal samples of 218 patients with chronic severe hepatitis B (CSHB) and 140 children with diarrhea were collected, and were examined for cryptosporidium oocytes by using auramine-phenol staining method (AA-p) and modified acid-fast staining method (MAF), and for cryptosporidium DNA by PCR and restriction digestion analysis. The factors affecting cryptosporidium infection of patients with CSHB were preliminarily analyzed. Results The positive rates of cryptosporidium infection detected by AA-p, MAF and PCR in the patients with CSHB and children with diarrhea were 4.1%, 3.2%, 6.0% and 0.7%, 0.7%, 1.4%, respectively. The positive rate of cryptosporidium infection detected by PCR in patients with CSHB was higher than that in children with diarrhea (P