Neelsen Method Research Articles

Urgent intraoperative cytological diagnosis of spherical mass lesions in the lungs

Differential diagnosis of pulmonary mass lesions is one of the most difficult tasks not only in pulmonology but also in phthisiology, because clinical manifestations, data from visual and laboratory examinations are undifferentiated in the early stages. In the case of an unclear diagnosis, morphological verification of the process played a crucial role. Urgent intraoperative examination is the most difficult option for morphologic verification of lung lesions, which requires a quick, detailed, and accurate answer.The aim of this study was to evaluate the clinical significance and efficacy of urgent intraoperative cytologic examination in the differential diagnosis between granulomatous (tuberculosis, sarcoidosis), tumor and tumor-like lesions of the lung.Methods. The results of urgent intraoperative cytologic examination of 173 tissue specimens from lung masses were retrospectively analyzed. Cytologic specimens were stained by the May – Grunwald – Giemse and Ziehl – Neelsen methods and then visualized on a Zeiss Lab A1 microscope (Carl Zeiss, Germany). The obtained data are compared with the results of histological examination. To evaluate the effectiveness of intraoperative cytologic examination, the accuracy, sensitivity, and specificity were calculated.Results. Analysis of the results of urgent cytological examinations (n = 173) shows that granulomatosis (tuberculosis/sarcoidosis) was established in 64.7% of cases, malignancy – in 31 cases (cancer – 9.2%, carcinoid tumor – 2.9%, malignant neoplasm of unclear etiology – 4%, suspected malignant lesion – 1.7%); benign process – in 9.2% of cases (hamartoma + chondroma – 5.2%, chronic inflammation – 4%). 1.2% of the cases had no diagnostic material. When comparing the obtained results, there was a discrepancy between the histological picture (2.9% of cases) and the primary clinical diagnosis (5.7% of cases). Sensitivity, specificity, and accuracy of cytologic examination of intraoperative material amounted to 95.9, 71, and 98%, respectively.Conclusion. The high sensitivity and accuracy (> 90%) of urgent cytologic diagnosis of pathologic processes in the lung under intraoperative conditions were demonstrated by the correct assessment of morphologic signs of a given lesion and by their comparison with the clinical picture and the results of additional examinations. Analysis of cytological specimens showed that urgent cytological examination significantly complements the morphological picture of lung lesions and can be considered as the first stage of phenotypic diagnosis.

The prevalence of the Cryptosporidium parvum among children in Ramadi city

The purpose of this study was to determine the prevalence of Cryptosporidium parvum among children in the city of Ramadi and made a comparison between ziehl -Neelsen method and the ELISA method, which is usually used to diagnose this parasite in Ramadi hospital (administration of Obstetrics and Gynecology Hospital and Ramadi Teaching Hospital).
 Fecal samples were collected from 813 children under 5 years. Oocysts of Cryptosporidium parvum parasite were determined by using ELSIA and Ziehl-Neelsen (acid-fast stain, ZN).
 Our study was shown that the rate of prevalence of the C. parvum was 7.87% from the total of 813 samples of diarrhea by using Zeihl-Neelsen method while ELISA technique result was 11.56% From our results, ELISA technique more specific than Ziehl-Neelsen method in diagnosis of C. parvum and the different is significant (P <0.05)..

الحدوث وعوامل الخطورة لطفيل الأبواغ الخفية بين مرضى نقص المناعة في مدينة إب – اليمن

Cryptosporidium parasite is causing opportunistic Cryptosporidiosis diseases among Immunocompromised patients that lead to acute and persistent diarrhea and health complications, especially in developing countries. This study amid to determine the occurrence of C. parvum among immunocompromised patients presenting at Al- Thawra hospital in Ibb city- Yemen. Sample of the study (87) stool specimens were collected between November 2016 and July 2017. The stool specimens were screened for the C. parvum by using formalin- ether concentration, Ziehl- Neelsen methods, and enzyme- linked immunosorbent assay (ELISA). The required data were obtained by a designed questionnaire. The result revealed that the overall rate of C. parvum was 62 (71.26%) observed among immunocompromised patients and both males and females had nearly equal frequency rates of infection. The highest rate of infection was 80% reported within age less than 10 years and 78.3% among patients who coming from the rural area. Also, it was found 83.33% of infected patients drinking from piped water with significant difference (P<0.05). In addition, 73.81% of individuals contacting animals were positive for infection. The most frequent rate of infection was 53.2% recorded for chronic renal failure, 42% for malignancy, and 4.8% for renal transplant patients. The clinical features most frequently presented were diarrhea, malaise, poor appetite, weight loss, abdominal pain, and fever. Similarly, 90.3% and 97.8% of cases were suffered from semi- liquid diarrhea and acute diarrhea, respectively. Recommendation as described in this study health education programs are needed for preventing and controlling the transmission of Cryptosporidium parasites among weak immune individuals.

Challenges in the morphological diagnostics of infectious lung diseases among HIV-infected patients: a case-series study

HIV-infection is a long-term infectious disease, characterized by deterioration of the immune system and it is often complicated by the development of opportunistic diseases, mainly of infectious origin. HIV-associated infections become generalized in the late stages of the disease and often may combine, that makes challenges in the diagnosis of these diseases. Differential morphological diagnosis of HIV-associated lung infections is quite complex. Most frequently, there is the need to differentiate between tuberculosis, bacterial or septic pneumonia, cytomegalovirus lung lesions, and deep mycoses. The aim. To demonstrate the expediency and necessity to apply special complex morphological methods in HIV-infected patients with secondary lung lesions using the series of clinical cases as an example. Materials and methods. The case series consisted of 7 cases with discordance between the clinical-radiologic, laboratory data and histological conclusions. Of these, there were 5 cases of life-time and 2 autopsy diagnoses. All the clinical and laboratory data of these cases were retrospectively analyzed, and a number of additional special histological examinations were performed. The Ziehl–Neelsen and fluorescent methods, Gomori–Grocot and alcian blue-PAS staining as well as immunohistochemical examinations were used for the detection of mycobacterial antigens with different antibodies to M. tuberculosis. Results. The case series of initial morphological misdiagnosis or indeterminate histological diagnosis was revealed among hospitalized HIV-infected patients. Clinical data did not provide an accurate diagnosis of pulmonary pathology. According to the traditional histological examination, a tuberculous process was diagnosed in 3 cases, a chronic granulomatous process – in 2 cases, but without specifying the etiology. In two cases of autopsy, pneumocystis pneumonia was suspected in combination with other infections. Additional morphological examination with using 2–6 special methods made it possible to determine the correct diagnosis. In 2 cases of provisional diagnosis of tuberculosis, invasive cryptococcosis and “pneumonia in a drug user” were revealed. Cryptococcosis and pneumocystis pneumonia were detected in 2 cases with an unspecified process. In the autopsy cases, the diagnosis of pneumocystis pneumonia was ruled out, while viral-bacterial pneumonia and tuberculosis associated with cytomegalovirus infection were detected. Conclusions. The presented factual material demonstrates the principal importance of using a comprehensive morphological examination in cases of lung inflammatory processes in HIV-infected patients. Clinical patient data, namely HIV-status at the time of diagnostic procedures as well as radiological and microbiological findings serve as a substantial aid in the diagnosis.

Histochemical characteristics and distribution of lipofuscin and polyglucosan bodies in the brain of dogs more than 10 years old

The ageing process is accompanied by numerous changes in the brain of dogs, such as accumulation of amyloid, fibrosis of blood vessel walls and meninges, accumulation of lipofuscin, and the presence of polyglucosan bodies (PGBs), satellitosis and neuronophagia. In this study, the presence of lipofuscin and PGBs in various parts of the brain in dogs of different sexes and ages was examined. For this purpose, brain samples were stained using haematoxylin eosin, modified Ziehl Neelsen and Periodic acid Schiff (PAS) methods. Lipofuscin was visualised by Ziehl Neelsen and PAS methods of specific staining on the same brain tissue segments. Lipofuscin had accumulated in 93% of old (more than 10 years old) dog brains, mostly in neurons of the medulla oblongata. The percentage of age-related lipofuscin pigment in other examined brain tissue segments was lower than in the medulla oblongata. There was a small difference in the percentage of lipofuscin-positive individuals between the two staining methods. The presence of PGBs was established by the PAS method for the vast majority (about 93%) of the old dogs (more than 10 years old), while PGBs were not detected in the group of young dogs (up to 5 years old). However, PGBs occurred in all examined segments of the dog?s brain tissues (for each of the tissue types, from 90% to 93% of dogs were positive for PGBs). The results obtained the oldest dogs (15 years old) harboured PGBs both extracellularly and intracellularly, while in other dogs, only extracellular PGBs were seen. Lipofuscin was accumulated mostly in large neurons of olivary nuclei of the medulla oblongata. PGBs were confirmed in all examined segments of the brain tissue of dogs more than 10 years old. This is one of the numerous indications that old dogs could be a very good animal model for studying the normal ageing process or neurodegenerative diseases.

Molecular detection of Rifampicin and Isoniazid resistance in culture isolates of newly diagnosed TB patients

Introduction: Multidrug-resistant tuberculosis (MDR-TB) is an emerging public health problem in many regions of the world, particularly in developing nations. Accurate and rapid diagnosis is essential in the management of MDR-TB, not only to optimize treatment but also to prevent transmission. Aims: To evaluate drug resistance in culture isolates by conventional and molecular methods and detect drug resistance gene in MDR-TB patients. Material and Method: 100 newly diagnosed pulmonary tuberculosis (TB) diagnosed patients attending TB Clinic, Gandhi Hospital, Secunderabad were included in the study. Two sputum samples collected from the patients were subjected to sputum microscopy, culture, Drug Susceptibility Testing (DST). Geno Type Mycobacterium Tuberculosis Drug Resistance (MTBDR) plus assay was done on the culture isolates to detect Rifampicin and Isoniazid (INH) resistance. Results: Out of 100 samples, 48 % smear positivity by Ziehl Neelsen (ZN) method, 51 % culture positivity on LJ medium,11.7% multi drug resistance for Rifampicin and Isoniazid with conventional drug susceptibility – Proportion method,17.6 % drug resistance by molecular method – Geno Type MTBDR plus was observed. Among the 4 Rifampicin (Rif) resistant isolates 2isolates showed mutation (mut) at D516V and in other 2 isolates only wild type (WT) was missing but no mut was seen . In the 1 Isoniazid (INH) resistant isolate WT was missing, but no mutation was seen. Among the 4 Rif +INH resistance all showed mut at S531L for RIF and at S315T1. Conclusion: The Genotype MTBDR assay is a rapid and reliable tool for the routine direct detection of MTB strains and of strains resistant to INH and RIF in smear positive, highly infectious patients. The rapid turn around time of the test enables the optimization of the therapy of these patients before confirmatory culture results are available. The test does not require viable organisms and thus reduces the biohazard risk in the laboratory.

Prevalence of Human Immunodeficiency Virus among tuberculosis patients in Kano, Nigeria

Background Tuberculosis (TB) is a major public health problem worldwide with an estimated one third of the world's population being infected by the agent Mycobacterium tuberculosis. The TB problem has escalated in the last two decades in the developing countries because of the recent pandemic of human immunodeficiency virus (HIV) infection. Previous studies have shown a high prevalence of HIV in TB patients. However, there is no documented study from Kano, Nigeria. Aims Therefore, this study is aimed at establishing current prevalence of HIV infection in TB patients attending Aminu Kano Teaching Hospital, Kano, Nigeria. Methods All confirmed pulmonary TB were screen for HIV. Pulmonary TB was diagnosed by microscopic examination of acid-alcohol fast bacilli stained sputum smear using the Ziel- Neelsen method. HIV was determined using Genscreen plus HIV Ag-Ab EUSA technique and confirmed by immunoconfirm technique. Results Among the 200 tuberculosis patients studied, 41(20.5%) were positive for antibodies to HIV. The highest prevalence rate was found in th age group 31 to 40 years (12%) Conclusion This study has confirmed high prevelance of HIV in patients with tuberculosis. The result shows a high HIV seroprevalence in TB patients in Kano, Nigeria. It is suggested that patients with TB should be tested for routinely for HIV.Key-words:Human Immunodeficiency Virus, Tuberculosis, Prevalence, Kano, Nigeria

Prevalence of Pulmonary Tuberculosis in Kanchanpur District of Far Western Nepal

Keeping in view of high prevalence of tuberculosis in far western development region in different reports, adoor to door survey of 125 household was carried out randomly in Mahendra Nagar municipality ward no13 of Kanchanpur district to pick up symptomatics suggestive of pulmonary tuberculosis and find the prevalence in the target population. A total of 200 sputum specimens from 70 symptomatics within the age rangeof 10-80 years were collected and subjected to smear microscopy by Ziehl - Neelsen's method. Out of 10positive cases (14.28%), infection in males and females was 80% and 20% respectively. The infection wasfound in the productive age group 20-69. The highest prevalence was obtained in 20-29 age group (25%)followed by 40-49 (11%), 30-39 (9%), 50-59 (8%) and 60-69 (8%). The diagnostic yield of single sputumspecimen examined was insufficient in field condition, especially where the sputum positivity was low. Of thethree sputum smears examined, the early morning specimen had the best result.Key Words: Pulmonary tuberculosis, prevalence, sputum smears.