Zeihl-Neelsen Staining Research Articles

Identification of Brucella spp. in Aborted Fetuses by Guineapig inoculation

Abstract Background: Brucellosis is a zoonotic disease which is endemic in Bangladesh. The prevalence of bovine brucellosis in subsistence management system in Bangladesh is low. However, the prevalence of bovine brucellosis at Central Cattle Breeding and Dairy Farm (CCBDF) is reported to be very high and Brucella abortus DNA has also been detected from cows’ milk of this farm. The principal manifestation of bovine brucellosis is abortion in pregnant cows, which is common at CCBDF. The role of brucellosis in abortion at CCBDF has not been ascertained. Hence, this study was undertaken to confirm Brucella spp. as the etiology of abortion in cows at CCBDF. Materials and Methods: Aborted fetal membranes and vaginal swabs from 3 cows, in which late abortion occurred, were collected aseptically from the CCBDF. The samples were initially stained with modified Zeihl-Neelsen staining method. The stain-positive samples were ground individually using a pestle and mortar, and a homogenized mixture was prepared by adding normal saline. Two milliliters of the homogenate from each sample were inoculated subcutaneously into a guinea pig. The sera of guinea pigs were collected after 3rd week of inoculation to perform rose Bengal test (RBT) and rapid antibody test (RAT). Result: Samples from two cows showed positive staining result in which numerous pink-colored coccobacilli were seen. All the sera collected were tested positive for both RBT and RAT. It is evident from this study that three of two abortions at CCBDF were due to brucellosis. Conclusion: Guineapig inoculation technique could be used as a good alternative of culture for confirming the diagnosis of brucellosis from contaminated clinical samples like placenta.

Genetic map of the isolated Cryptosporidium parasite from children with diarrheal in the city of Mosul

The current study was conducted between the beginning of October 2022 and the end of March 2023 to identify the prevalence of Cryptosporidium parivum infection among children under five years using microscopic and molecular methods. The study recorded 43(37.71%) infected children diagnosed using the Modified Zeihl Neelsen Stain and Gimsa stain. On the other hand, a molecular examination of stool samples using PCR showed that 25(21.92%) children out of 115 collected stool samples were infected with this parasite. DNA Sequencing was performed for specimens diagnosed with Cryptosporidium parasite, and the new isolates were registered for the first time in Nineveh governorate in NCBI under the accession number 757376. Keywords: Cryptosporidium disease, Genetic map.

Cutaneous tuberculosis presenting as abdominal wall cellulitis: a case report

Tuberculosis (TB) continues to be a prevalent disease worldwide; according to World Health Organization (WHO), children and young adolescents represent about 11% of all people with TB globally. Although TB often is cited as a disease that commonly occurs in underdeveloped countries, the evolution of drug-resistant forms of TB and infection sensitivity of immunocompromised individuals have made this disease as a focal point for developed countries as well. Although pulmonary TB is the most common form worldwide, but TB can infect any organ of the body. An uncommon version- cutaneous TB- affects <2% of all individuals with active form of TB. This case report describes a 4-month-old child who presented to us with abdominal wall cellulitis following a history of fall related injury. Incision and drainage of the pus from the fluctuant area grew acid fast bacilli on Zeihl-Neelsen staining. After the child didn’t show any improvement with intravenous antibiotics, empirical anti-tuberculosis therapy was initiated after which there was marked improvement. Cutaneous TB constitutes a very small percentage of extra-pulmonary tuberculosis and is rarely seen in paediatric age group.

Is nucleic acid amplification test sufficient enough for reducing the diagnostic dilemma of cervical lymph node tuberculosis? A descriptive study in a tertiary care hospital

Background: Pulmonary tuberculosis (TB) is common clinical presentation in India, but, nowadays, extrapulmonary TB (EPTB) is also a clinical problem. At present, no single test is available for diagnosing EPTB which is rapid, cost-effective, and confirmatory. Many studies were done to diagnose and treat the pulmonary TB, though more efforts are needed for diagnosis of EPTB. Aims and Objectives: The aim of this study is to find out any test or a series of tests that reduce the diagnostic dilemma of cervical lymph node TB. Materials and Methods: A total of 170 patients with cervical lymphadenopathy were enrolled in the pathology department between July 1, 2021, and December 31, 2021. It was a descriptive study and of cross-sectional design. The Institutional Ethics Committee approval was received (No. BSMC/IEC/101 dt. January 15, 2021). Suspected malignancy cases were excluded from the study. Fine-needle aspiration (FNA) was done for all these patients and stained with Leishman-Geimsa, Papaniculaou, and Zeihl-Neelsen (ZN) stain. After FNA, all samples were sent for cartridge based nucleic acid amplification test. Results: After FNA cytology, ZN stain was done and only 9.2% of acid–fast bacilli (AFB) positive cases were found. Granuloma was found in 27.6% of cases. After nucleic acid amplification test (NAAT) study, it was found that 36.8% patients were positive. Total number of lymph node considered to be positive for TB (NAAT positive and/or ZN stain positive and/or presence of granuloma) is 85 (52.1%). As per conventional method for diagnosis of lymph node TB (presence of granuloma and/or AFB positive in ZN stain), number of cases were 50 (30.7%). NAAT test was significantly more positive in respect of ZN staining but not significantly positive in respect of presence of granuloma. Conclusion: For diagnosis of cervical lymph node TB, all three methods (NAAT, ZN stain, and Leishman-Giemsa stain) should be used simultaneously.

Molecular Characterisation of Cryptosporidium parvum among Cattle and Cattle Handlers from Tripura (India) and the Associated Risk Factors

Background: Cryptosporidium parvum is major protozoan parasite of both animals and humans with zoonotic significance. There is a paucity of information on its occurrence and associated risk factors in North Eastern states, India. The present study aimed at the molecular characterization of Cryptosporidium from cattle and cattle handlers in West district of Tripura and risk associates analysis. Methods: Faecal samples were randomly collected from cattle and cattle handlers (100 each), of West district, Tripura. The samples were subjected to sheather’s sucrose flotation, modified Zeihl Neelsen (mZN) staining and PCR assay targeting the 18S SSUr-RNA gene for detection of Cryptosporidium. The polemerised chain reaction- Restriction length polymorphism (PCR-RFLP) was done to detect Cryptosporidium spp. and C. parvum genotypes. Association of the epidemiological variables in relation to host and environment were studied for comparative analysis. Result: The incidence of Cryptosporidium was 16 per cent and 7 per cent in cattle and cattle handlers, respectively. The molecular characterisation based on PCR-RFLP analysis revealed 14 C. parvum and two C. andersoni and three C. parvum in cattle handlers. All the 14 C. parvum from cattle and one C. parvum from cattle handlers belonged to bovine genotype (genotype II) and other two C. parvum from cattle handlers belonged to the human genotype (genotype I). Cryptosporidiosis in both cattle and cattle handlers were strongly associated with younger age group and diarrhoeic faecal consistency with significantly (p less than 0.01) higher occurrence. However, significantly (p less than 0.05) higher predominance of monsoon season in occurrence of the disease was observed in cattle and sex predominance was not observed in both cattle and cattle handlers.

Utility of cartridge based nucleic acid amplification test in diagnosis of tubercular lymph nodes on fine needle aspiration and its comparison with zeihl-neelsen staining and concentration bleach method

Tuberculosis (TB) is one of the oldest disease known to humanity and among the deadliest epidemics and is in the top 10 causes of death worldwide. The incidence of tuberculosis in India has dramatically increased and contributing to it is Human Immunodeficiency Virus (HIV). Various diagnostic methods are available for detection of tubercular bacilli. Only full proof criteria for diagnosis of TB is to demonstrate the bacilli from affected tissue. This study was planned in extra pulmonary tuberculosis cases to evaluate different staining methods for demonstration of mycobacterium on FNAC material. To evaluate efficacy of Cartridge Based Nucleic Acid Amplification Test (CBNAAT) in the initial cytodiagnosis of Tubercular Lymphadenitis 147 clinically suspected cases of extra pulmonary tuberculosis mainly TB lymphadenitis who attended cytology OPD were selected irrespective of age and sex. FNAC was performed. Slides stained for PAP, ZN and concentration bleach. Material remained in hub was sent for CBNAAT. PAP stained slides were observed for various morphological pattern.Six morphological patterns were observed. Positivity for acid fast bacilli by ZN staining, concentration bleach method and CBNAAT were compared. Detection of acid-fast bacilli by CBNATT was found more as compared to ZN and Concentration bleach method.

ROLE OF FIBEROPTIC BRONCHOSCOPY IN SPUTUM NEGATIVE PATIENTS

Background:Tuberculosis continues to be the one of the most common global health problems for more than a century. Fiberoptic bronchoscopy may be very useful in diagnosing cases who have no sputum or whose sputum smear is negative for Acid-Fast bacilli. The current study aims to assess the role of fiberoptic bronchoscopy in sputum smear negative under NTEP and radiologically suspected cases of pulmonary tuberculosis. Methods:This study was conducted in our institution in clinico-radiological suspected cases of pulmonary tuberculosis patients in whom two sputum smear for acid fast bacilli by Zeihl-Neelsen stain under NTEP was negative. Fiberoptic bronchoscopy was performed in all these patients and samples taken were sent for investigations. Result:Fiberoptic bronchoscopy was performed in a total of 110 patients of presumptive pulmonary tuberculosis whose sputum for AFB smear was negative. Out of 110 patients, 65.45% were male and 35.55% were female. Fever was the most common presenting symptom and was found in 93.64% patients. Radiographically, pulmonary consolidation was bilateral in 63.34% cases while was unilateral in 36.36% cases. The total yield of FOB came out to be 59.1% in which BAL for AFB smear was positive in 43.63% cases, BAL for Mtb culture was positive in 54.54% cases while, BAL for PCR was found to be positive in 59.09% cases. Conclusion:Fiberoptic bronchoscopy may be useful in rapid diagnosis and early treatment initiation thereby, reducing the risk of transmission of the disease. The findings of the study indicated that fiberoptic bronchoscopy is a useful method to diagnose the cases of sputum negative radiologically suspected symptomatic patients of tuberculosis.

Study on role of cytology in diagnosis of head and neck lesions in a tertiary care hospital

Background: Head and neck lesions are commonly encountered in patients across all age groups. This region encompasses a multitude of congenital, inflammatory or neoplastic lesions. FNAC is a clinically appropriate procedure associated with minimal trauma and complications for patients for analysis of head and neck swellings because of factors such as easy approachability of the target site, increased compliance and helping to avoid surgery in non-neoplastic lesions, inflammatory conditions and also some tumors. Objective: This study is undertaken to assess the accuracy of cytopathological diagnosis of head and neck lesions by FNAC and to correlate and analyze FNAC of head and neck lesions with that of histopathological study. Material and Methods: The present study was Laboratory based Cross sectional descriptive type of observational study. A total of 100 cases undergoing FNAC among the head and neck lesion were enrolled for present study. FNAC was done using 10 cc disposable syringe and 22/23 gauge needle taking all aseptic precautions. Both aspiration and non-aspiration techniques were used wherever required. Smears were stained with PAP, Haematoxylin-Eosin stain and Leishman stain. Zeihl-Neelsen staining for acid fast bacilli was done in suspected tubercular lesions. Results: Out of 100 patients lymph node lesions (46%) was the predominant site of FNAC. Out of 46 lesions of Lymph node; tubercular lymphadenitis was the most common pathological findings. Among 28 thyroid lesion Colloid goitre (45.5%) was the predominant finding. Out of 18 salivary gland lesions the most common lesion was pleomorphic adenoma. Out of 14 soft tissue and miscellaneous lesions benign lesions were commonest finding. Conclusion: FNAC is a simple, rapid and cheap diagnostic tool that can be done as an outpatient procedure with overall accuracy rate of more than 90 % which differentiates non-neoplastic lesions from neoplastic lesions thus avoiding unnecessary surgeries.

Intestinal parasitic infections among hemodialysis Sudanese patients.

Background: Intestinal parasitic infections (IPIs) can result in high morbidity and mortality, particularly in immunocompromised patients. Intestinal parasitic infections rate among hemodialysis Sudanese patients studied infrequently. Our paper aimed to examine and determine the prevalence of intestinal parasites among hemodialysis Sudanese patients. Methods: Stool samples were collected from Sudanese patients at a hemodialysis center in Kosti city, White Nile state, Sudan. All stool samples were examined to detect the type of parasites and determine the prevalence using the formol-ether concentration technique and Modified Zeihl Neelsen (ZN) stain technique. Results: The overall prevalence of Intestinal parasitic infections among hemodialysis using the formol-ether concentration technique and Modified Zeihl Neelsen (ZN) stain technique was (28.6%) and (44.6%) respectively. The total prevalence of parasites detected by formol-ether concentration technique; Giardia lamblia (46.9%), Entamoeba histolytica (12.5%), Entamoeba coli (40.6%), and by modified ZN staining technique; Cryptosporidium species (28%) and Isospora belli (8%). The prevalence of IPIs was higher among males than females (74 vs. 26%, respectively; P>0.05) Conclusions: Studying the prevalence of IPIs among hemodialysis Sudanese patients, may predict their health status and lead to better diagnosis and treatment. Health education and personal hygiene were recommended to reduce the transmission of parasitic diseases. This is a first descriptive study of IPIs among hemodialysis in the area of study.

Acid and heat fastness in microsporidia: How acid fast are acid fast microsporidium?

Microsporidia are obligate spore-forming microorganisms with strong resemblance to fungi and can affect almost every organ system in immunocompetent or immunocompromised individuals. Mixed infections are also reported in immunocompromised hosts. Microsporidial spores show marked morphological variations and the small and slender forms can resemble bacilli. Modified Zeihl Neelsen (ZN) stain, cold method demonstrates them as bright red in color, leaving several spores blue or incompletely stained; thus, they are reported as weakly or variably acid fast. Variability in staining results with ZN stain and considering the fact that Mycobacterium tuberculosis, the commoner bug in developing countries is identified by its resistance to stronger acids on ZN staining, authors wished to demonstrate acid and heat fastness in microsporidium using corneal tissue specimens. Microsporidial spores stained bright red in color with conventional ZN stain, demonstrated strong acid fastness, and interestingly the staining results improved on heating. Thus, the authors conclude that they are strongly acid and heat fast and care must be warranted so that they are not misdiagnosed as Mycobacterium or other acid-fast organisms. Careful observation of morphology, battery of special stains, and molecular diagnostics should be advocated for diagnostic confirmation. To the best of the authors' knowledge, this is the first explicit report on acid and heat fastness on microsporidial spores.

Cryptosporidiosis And Other Intestinal Parasitic Infections And Concomitant Threats Among HIV-Infected Children In Southern Ethiopia Receiving First-Line Antiretroviral Therapy.

BackgroundChildren infected with human immunodeficiency virus (HIV) are at high risk of acquiring intestinal parasitic infections. This study aimed to determine the magnitude of Cryptosporidium and other intestinal parasitic infections and concomitant threats among HIV-infected children.MethodsA hospital-based cross-sectional study was carried out at three antiretroviral therapy clinics in southern Ethiopia from February 2016 to June 2017 in 384 HIV positive children. Socio-demographic and clinical data were collected using structured questionnaires. Direct stool microscopic examination and modified Zeihl–Neelsen staining technique to identify parasites. Chi-square test was conducted to determine the real predictors of the infection. Significant association was considered when p-value <0.05 at 95% CI.ResultsThe overall magnitude of intestinal parasitic infections among the study population was 16.9% (95% CI: 13.0–20.8%). The most predominant parasitic infections were Cryptosporidium spp. (9.6%) and the least was Taenia spp. (0.78%). Diarrheal status (χ2=7.653, df=2, p=0.022) was detected to be the only significant associated variable.ConclusionCryptosporidium infection was found to be the most common intestinal parasitosis among HIV-infected children. Routine screening service for Cryptosporidium and other intestinal parasites is important in the clinical management of HIV-infected children.

Tubercular Lymphadenitis Mimicking Findings of Metastatic Signet Cell Carcinoma in FNAC: A Case Report

Tubercular lymphadenitis is one of the most common extrapulmonary tubercular lesions presented in the Outpatient Department which is commonly diagnosed by Fine needle aspiration cytology. Cytological diagnosis of tuberculosis requires the presence of epithelioid cell granulomas demonstration with or without Langhan’s giant cells and necrosis. Sometimes, there can a proliferation of signet like cells without granulomas, which in a cervical lymph node can be given a false interpretation of metastatic signet cell carcinoma.&#x0D; A 35 year female patient with a complaint of cervical lymphadenopathy came to the medical OPD. FNAC was done which showed numerous scattered signet ring cells. However, epithelioid cell granulomas weren’t observed in the smears. Zeihl Neelsen stain for Acid fast bacilli was done but the organism wasn’t appreciated. A biopsy was done and a histopathological slide showed epithelioid cell granulomas and Langhan’s giant cell. Zeihl Neelsen Stain for Acid fast bacilli, which was positive.

A cytological study of smear characteristics and its correlation with bacillary load in tubercular lymphadenopathy

Abstract Introduction: Extrapulmonary tuberculosis in form of lymphonodes is the common presentation of tuberculosis in developing countries. Fine needle aspiration cytology helps in early diagnosis & treatment for tubercular lymphadenitis, which remains the cornerstone for the effective management of the disease. Aim: To study the cytological characteristics and compare with the bacteriological load with the help of Zeihl Neelsen stain. Materials and Methods: 445 smears were studied for a period of one year. Smears were categorized into 4 patterns Pattern A - epithelioid cell granulomas without caseous necrosis, Pattern B -epithelioid cell granulomas with caseous necrosis, Pattern C- caseous necrosis without granuloma, Pattern D – neutrophils with abundant caseous necrosis. Bacillary load was graded as 1+, 2+ and 3+ were compared with the cytological categories. A p value of Results: Pattern A was observed in 16.7% cases, Pattern B in 18.3%, Pattern C in 37.8% & Pattern D in 27.2% of cases. Statistical significance was seen between cytomorphological pattern & AFB grade (p=0.026 & X2=20.0639). It was found that pattern C& D showed the highest AFB grade of 3+ by a 2x2 analysis to test the degree of one hypothesis. Conclusion: FNAC is an useful tool in the early diagnosis of tubercular lymphadenitis avoiding the un necessary burden of surgical intervention. Maximum bacillary load was correlated with the patterns C & D. Keywords: Fine needle aspiration cytology, Tuberculosis, Lymph nodes, Caseous necrosis, Acid fast bacilli. Introduction Tuberculosis is a major public health problem worldwide, especially in the developing countries like India. Peripheral lymphadenopathy is the most common manifestation of extrapulmonary tuberculosis.[1],[2]Fine Needle Aspiration Cytology (FNAC) is safe, cost effective and conclusive procedure.[3] It provides an alternative to excision biopsy for lymph nodes and is an easy procedure for collection of

Evaluation of GeneXpert MTB/RIF assay in diagnosis of extrapulmonary tuberculosis and rifampicin resistance

Background: The prevalence of tuberculosis is still on the rise particularly extrapulmonary cases. There is an urgent need for rapid diagnosis of these cases for prompt initiation of treatment. Aim: To evaluate the role of GeneXpert MTB/RIF Assay in the diagnosis of clinically suspected extrapulmonary tuberculosis (EPTB) and to detect rifampicin resistance in these cases. Materials and methods: A total of 241 samples from April 2016 to July 2018, from all clinically suspected EPTB patients with support of either laboratory or radiological evidence were included in study. 132 pleural fluid, 59 lymph node aspirate, 21 CSF, 17 pus, 9 ascitic fluid and 3 synovial fluid samples were screened for presence of acid fast bacilli (AFB) by conventional Zeihl-Neelsen (ZN) technique. The same samples were also used for testing by GeneXpert MTB/RIF Assay. Results: Out of 241 EPTB samples tested, none were positive for AFB by ZN staining. Overall 9.54 % (23 out of 241) samples were positive for MTB by GeneXpert with a positivity rate of 23.80%, 13.55%, 11.76% & 6.06% respectively for CSF, lymph node aspirate, pus and pleural fluid. MTB was not detected in any of ascitic fluid & synovial fluid samples. Of the 23 MTB positive samples detected by GeneXpert, all were sensitive to rifampicin. Conclusions: GeneXpert was found to be simple, effective and useful diagnostic method for detection of EPTB due to its rapidity & simultaneous detection of rifampicin resistance, thus reducing the time taken for initiation of treatment.

INCIDENCE AND EXPERIMENTAL INFECTION OF CRYPTOSPORIDIUM BAILEYI IN CHICKEN

This study was aimed to investigate the incidence of Cryptosporidiosis in 200 fecal samples from slaughtered broiler chicken carcasses in the local markets in some areas of Baghdad city (Al-Hurriya, Al -Kadhimiya and Al-Shuala), during March to May 2017. Three diagnostic techniques used (flotation by Sheather's sugar solution, staining with Modified Zeihl-Neelsen stain, and measuring of isolated Cryptosporidium oocysts by ocular micrometer) to determine the type of Cryptosporidium species, and for conform that the isolated species of parasite from infected cases belong to the C.baileyi. Experimental infection done in, 18 broiler chicken chicks aged one week divided to three groups first (G1) and second (G2) inoculated orally with, (500, 1000) oocysts per chick respectively, while the third group remain as a control (G3), than pathological lesions detected in some internal organs of infected chicks (trachea, intestine and bursa of fabricius). The study recorded a total infection rate 35% (75/200) in slaughtered broiler chicken. The result were revealed that the highest rate of infection occurs in April, reached 46% (23/50), while the lowest rate of infection in June, reached 20% (10/50). The experimental study revealed, the infection was occur in all chicks, of (G1) and (G2), and the first clinical signs appear after 7days post infection (PI) which include diarrhea, dullness, anorexia, and increased consumption of water, which represented the incubation period of the parasite, while the shedding of oocysts in feces started after 6-9days PI.

Zeihl Neelsen Stain: Still a Reliable Option for EPTB Diagnosis in Resource Constraint Settings

Background: Endemic countries like India are burdened with the bludgeoning effects of tuberculosis. While pulmonary tuberculosis is more prevalent and communicable, diagnosis of extra pulmonary tuberculosis (EPTB) is more challenging.Methods: This was a retrospective study done for three consecutive years 2013 to 2015. The extra-pulmonary smear samples from patients suspected of tuberculosis were stained with Zeihl Neelsen stain and examined. Results: Based on ZN stain, EPTB was diagnosed in 13.4%, 9.5%, 10.6% suspected cases between 2013 to 2015 respectively. Highest positivity was seen among pus aspirates from lymph nodes in 13-25 years age group.Conclusion: New technological advances are being made all across the globe for faster and specific tuberculosis diagnosis. However in resource limited settings, reliance is often on the smear microscopy findings. Diligently observed smears increase chances of diagnosis and is often rewarding.

Diagnostic Potential of Circulating Biomarkers in Adenosine Deaminase Diagnosed Pleural Tuberculosis Cases.

Pleural tuberculosis accounts for nearly 20% of Extra pulmonary tuberculosis. Adenosine deaminase, commonly used biomarker for the diagnosis, is non specific and there is paucity of literature on its correlation with conventional or newer methods for the diagnosis of extra pulmonary forms of TB. The aim of the study was to assess diagnostic potential of T cell function markers [interferon (IFN-γ), interleukin (IL-2) and IFN-γ/IL-2 ratio]; macrophage activation marker [neopterin]; and oxidative stress markers [protein carbonyl and malondialdehyde (MDA)] in pleural tuberculosis. 26 pleural TB cases diagnosed on the basis of suggestive chest X-ray and raised serum ADA levels and healthy controls were included in the study. Pleural fluid specimens were subjected to Zeihl Neelsen staining and culture on Lowenstein Jensen medium. Serum IFN-γ, IL-2, neopterin and protein carbonyl levels detection were done by ELISA and MDA levels were determined by measuring the thiobarbituric acid reactive substances. Median serum levels of IFN-γ, IL-2, IFN-γ/IL-2 ratio, neopterin, protein carbonyl and MDA were significantly different between cases and controls. Levels of all biomarkers except IL-2 were significantly higher in cases with contact history. Mean levels of ADA and ESR were 46.27U/L and 46.62mm/hr in PTB cases. AUC for IFN-γ, IL-2, IFN-γ/IL-2 ratio, neopterin, protein carbonyl and MDA were significantly discriminative for cases and controls. IFN-γ/IL-2 ratio was best discriminatory biomarker with highest area under ROC curve. Though no correlation was seen between ADA and any of the six biomarkers, ESR levels correlated significantly with all biomarkers except IL-2 by spearman's correlation coefficient. Though all the circulating biomarkers under study provide useful supportive evidence for the diagnosis of PTB, further studies involving diverse control groups particularly non-PTB effusion are needed to validate these results.

Utility of PCR targeting IS6110 and MPT64 genes in the diagnosis of extrapulmonary tuberculosis

Abstract Introduction Extrapulmonary tuberculosis (EPTB) is one of the major diagnostic challenges in clinical practice. The aim of the study was to compare the utility of Polymerase Chain Reaction (PCR) with conventional techniques in the diagnosis of EPTB. Materials and methods Specimens from 150 patients with a presumptive diagnosis of EPTB were examined by Zeihl-Neelsen staining and mycobacterial culture on Lowenstein Jensen medium. Histopathology was done for lymph node aspirates and endometrial biopsies. DNA from all the samples was amplified by PCR targeting 123 bp and 240 bp fragments of IS6110 and MPT64 genes respectively. Results IS6110 PCR was more sensitive (82.1%) as compared to PCR for MPT64 gene (81%) though the difference was not found to be statistically significant. Four (2.6%) and 2 (1.3%) samples were positive by IS6110-PCR only and MPT64-PCR only respectively.Conclusion: PCR with multiple targets can have potentially important role in improving diagnostic accuracy of EPTB.

Front loading sputum microscopy – an alternative approach for diagnosis of pulmonary tuberculosis

IntroductionUntil newer, rapid, economical tools are introduced for diagnosis of Pulmonary Tuberculosis in resource limited settings, optimization of sputum smear examination for increasing case detection remains of utmost priority. The aim of the study was to detect presumptive TB patients using Front Loading sputum microscopy and compare it with Standard method. MethodsThree sputum specimens (Spot 1- on spot at the time of first visit, Spot 2- one hour after Spot 1 and early morning-next day early morning sample) from 552TB suspect cases were collected. Zeihl Neelsen staining (spot 1, spot 2 and early morning respectively) and microscopy by Front Loading (spot 1, spot 2) and Standard method (spot 1, early morning) of sputum microscopy were done. ResultsCulture on LJ media being the gold standard, the sensitivity and specificity of the Front Loading and the Standard method of sputum microscopy were 68.65%, 94.43% and 70.14%, 93.6% respectively. The difference between two methods was not statistically significant. 91.1% patients gave preference for same day sampling process. ConclusionThe sensitivity and specificity of sputum microscopy using an early morning sample followed by another sputum one hour later from the same day appears not to be inferior to using two early morning samples on subsequent days. The Front Loading sputum microscopy can be implemented in DOTS clinic on the day of first visit of patients to health care center to increase compliance of patients with diagnostic procedure and decrease drop-outs.

Clinical and cytological features in diagnosis of peripheral tubercular lymphadenitis – A hospital-based study from central India

BackgroundTuberculosis lymphadenitis is difficult to diagnose clinically, and often the laboratory confirmation is not available in resource-poor countries. We describe here the symptoms, clinical characteristics, and results of cytological analysis in peripheral tuberculous lymphadenitis patients. MethodsOne hundred and fifty-six patients with peripheral lymph node for cytological evaluation presenting to Department of Pathology, Acharya Vinoba Bhave Rural Hospital, Wardha, India were included in this study. ResultsSixty-nine cases were tuberculous lymphadenitis, with female to male ratio of 1.3:1. One or more constitutional symptoms were present in 59.4% of patients, with 89.9% of lymph nodes ≥2×2cm and the most common site of involvement was cervical lymph node (70.3%). The lymph nodes were multiple (85.5%), either discrete or matted. Cytomorphologically, hemorrhagic aspirate was observed in 29 cases, well-formed epithelioid cell granuloma with caseous necrosis was seen in 34 cases, and Zeihl Neelsen staining was positive in 45 cases. Correlation between character of aspirate and cytomorphological pattern was found highly significant. ConclusionThese data suggest that constitutional symptoms and clinical and cytological features help in diagnosing cases of peripheral tubercular lymphadenitis and also open new frontiers to further research that affects the cytological features of these cases.