Detection Of Acid-fast Bacilli Research Articles

Cytomorphological findings in confirmed cases of tubercular lymphadenitis.

Tuberculosis (TB) remains a major health problem, especially in the developing countries. Fine-needle aspiration cytology is the first line of investigation for tubercular lymphadenitis as it is easy to perform, less invasive, quick, and economical. The typical cytopathological features of tuberculosis TB include epithelioid cell granulomas with Langhans giant cells and caseous necrosis. The present study aimed to evaluate the cytomorphological features of newly diagnosed cases of tubercular lymphadenitis confirmed by GeneXpert. This was a retrospective study in which all fine-needle aspirates from newly diagnosed cases of tubercular lymphadenitis confirmed by GeneXpert over a 1-year period from July 2022 to July 2023 were included in the study. The May-Grunwald-Giemsa stained smears from these aspirates were categorized into three patterns-epithelioid cell granulomas with necrosis, epithelioid granulomas without necrosis, and necrosis only. The granulomas were further categorized into well-formed, ill--formed, and splintered. The background of the aspirate which included a reactive lymphoid background, lymphocytes, eosinophils, and neutrophils was tabulated for all the cases. Out of the three cytomorphological patterns, epithelioid granulomas with necrosis were the most predominant (67.5%), followed by necrosis only (20.8%) and granulomas without necrosis (11.6%). An acid-fast bacilli (AFB) positivity of 53.3% (64 cases) was seen on the Ziehl-Neelsen stain. Well-composed, poorly formed, and splintered granulomas were seen in 55 (57.9%), 42 (44.2%), and 21 (22.1%) cases, respectively. Neutrophils were the most common background population (42, 35%) followed by lymphocytes (36, 30%). Reactive lymphoid cells and eosinophils were seen in 12 (10%) and 3 (2.5%) cases, respectively. Fine-needle aspiration cytology is a rapid inexpensive minimally invasive test for tubercular lymphadenitis as epithelioid cell granulomas along with caseous necrosis are highly suggestive of TB. However, manual acid-fast AFB detection has a low sensitivity as is illustrated in the present study where only AFBwas demonstrated in only 53.3% of cases.

Prevalence of pulmonary tuberculosis and HIV infections and risk factors associated to tuberculosis in detained persons in Antananarivo, Madagascar

The incidence rate of tuberculosis in prisons is estimated to be 8 times greater than that in the general population in Madagascar. Our objectives were to estimate the prevalence of pulmonary tuberculosis and HIV infection among prisoners and to identify risk factors associated with tuberculosis. We conducted a cross-sectional study at the central prison of Antananarivo from March to July 2021. Individual male and female inmates aged ≥ 13 years who had lived in the prison for at least three months prior to the study period were included as participants. Acid-fast bacilli detection by microscopy and/or culture, an intradermal tuberculin test, a chest X-ray, and a rapid diagnostic orientation test for HIV were performed. Among 748 participants, 4 (0.5%) were confirmed to have pulmonary tuberculosis. Overall, 14 (1.9%) patients had “confirmed” or “probable” tuberculosis [0.90–2.84, 95% CI]. The proportion of participants with latent tuberculosis infection was 69.6% (517/743) based on a positive tuberculin test without clinical symptoms or radiography images indicating tuberculosis. Out of 745 HIV screening tests, three showed reactive results (0.4%). Age (OR = 4.4, 95% CI [1.4–14.0]) and prior tuberculosis treatment (or episodes) were found to be associated with confirmed and probable tuberculosis.

Utility of Conventional ZN Staining and Modified Bleach Techniques in Detection of Acid Fast Bacilli from Aspirates of Suspected Cases of Tuberculous Lymphadenitis

Introduction: Tuberculosis (TB) is a significant public health problem in many developing countries, including India. Diagnosing tuberculous lymphadenitis, in particular, poses challenges as it requires the demonstration of Acid-Fast Bacilli (AFB) using the conventional Ziehl-Neelsen (ZN) technique. The Modified Bleach Technique, which involves liquefying the aspirated specimen with sodium hypochlorite followed by centrifugation, has been described to enhance the yield of AFB, improving sensitivity and diagnostic accuracy. Aim: To highlight the role of the bleach concentration method compared to conventional ZN staining in detecting AFB in Fine-Needle Aspiration (FNA) material from clinically suspected cases of tuberculous lymphadenitis. Materials and Methods: A prospective cross-sectional study was conducted at the Department of Pathology, MVJ Medical College and Research Hospital, Bengaluru, Karnataka, India from November 2020 to December 2022. Fine needle aspiration material from 100 cases of clinically suspected tuberculous lymphadenitis was included. In each case, part of the aspirate was processed for ZN staining, while the remaining material was used for modified bleach staining and Cartridge Based Nucleic Acid Amplification Test (CBNAAT). Patient data, including age, sex, duration and site of swelling, nature of aspirate, and cytomorphological diagnosis, were documented. The data was entered into a Microsoft excel spreadsheet and analysed using the Chi-square test, with a p-value<0.05 considered statistically significant. Results: The age range of the study participants was 1-80 years. Out of 100 cases, 13 were excluded due to inadequate material. The most common lymph node site was cervical 69 (79.3%), followed by axillary 8 (9.1%) and supraclavicular 6 (6.89%). Among the 87 cases, reactive lymphadenitis was diagnosed in 36 (41.3%), suppurative lymphadenitis in 12 (13.8%), necrotising granulomatous lymphadenitis in 12 (13.8%), granulomatous lymphadenitis in 15 (17.3%), and necrotising lymphadenitis in 12 (13.8%). Out of the 100 cases included in the study, 32 cases were confirmed as tubercular lymphadenitis by CBNAAT. Among these 32 cases, conventional ZN stain was positive for AFB in only 3 cases (9%), whereas the Modified bleach stain was positive in 16 cases (50%). The results were compared with CBNAAT findings, and the modified bleach technique showed sensitivity (59.4%), specificity (100%), Positive Predictive Value (PPV) (100%), Negative Predictive Value (NPV) (80%), and Accuracy (85%). Conclusion: The Modified bleach method of ZN staining is a simple, safe, inexpensive, and easy-to-perform technique. With the bleach method, AFB was easily visible and detectable in the majority of AFB-positive cases. Additionally, the bleach method preserved the morphology of AFB better. It improves the microscopic detection of AFB, leading to a definitive diagnosis of TB on cytology.

Comparision of pap stained smears by using auto fluorescence method with ziehl-neelsen method for detection of acid fast bacilli (AFB)

The Ziehl-Neelsen staining method though it plays an important role in detecting acid fast bacilli (AFB) by direct microscopy its low sensitivity makes to look for the other alternatives. : This study was undertaken to evaluate the utility of auto fluorescence method on Papanicolaou stain (Pap) in comparison tothe traditional Ziehl-Neelsen stain (ZN) method in detection of AFB from lymph node aspirates. Fine needle aspirations (FNAs) were done in 153 patients with clinical suspicion of tubercular lymphadenitis. Smears from aspirate were processed for routine cytology for Hematoxyllin & Eosin (H&E), Giemsa, Pap, Ziehl-Neelsen (ZN) and Auramine (AR) staining. Pap stained smears were examined for AFB for their autofluorescence under fluorescent microscope using the blue excitation filter. ZN stained smears are examined for AFB under oil immersion of compound microscope. The efficacy of the autofluorescence method by Pap in detection of acid fast bacilli (AFB) over the conventional Ziehl-Neelsen stain was analyzed by taking AR as a standard control for AFB.Tuberculous lymphadenitis was diagnosed in 36 cases out of 153 clinicall suspected cases by cytology, culture and microscopyThe sensitivity of ZN was 57.14% and specificity was 99.15% while the sensitivity of AF was 96.5% and the specificity was 91.8%. : It was a novel method of detection of AFB, cheaper, easily available and less time consuming than other methods. AR also had high sensitivity rates as AF.

Pulmonary tuberculoma: does persistent imaging lesion equals treatment failure?

Pulmonary tuberculoma is a rare but challenging entity for paediatricians, regarding diagnosis and management. Treatment course is still up for discussion since tuberculomas may persist after treatment. The authors report two paediatric cases, both recently treated for latent tuberculosis infection, admitted for investigation of a solid pulmonary nodule. In both cases the thoracic CT scan showed a nodular subpleural lesion and the biopsy revealed epithelioid cells with areas of necrosis and calcification. The diagnosis of pulmonary tuberculosis was confirmed in one case by detection of acid-fast bacilli in the smear and in the other by nucleic acid amplification test for M. tuberculosis. The patients presented good response to treatment with antituberculosis drugs, without need for surgical resection. These reports highlight the diagnostic and treatment challenges of this rare entity.

An Interesting Case Report on Calcaneal Tuberculosis as a Rare Presentation of Disseminated Tuberculosis

Despite the high prevalence rate of tuberculosis infection in Malaysia, calcaneal tuberculosis (TB) is rare. In this article, we present a case of a 53-year-old Malay female who initially presented with left calcaneal swelling for 3 months. Surgical drainage was performed and affected calcaneal was biopsied for histological examination which findings reported as consistent with tuberculosis infection. Further history revealed concomitant respiratory symptoms with the diagnosis of disseminated TB is supported by positive Mantoux test, detection of Acid-Fast bacilli in sputum and CT imaging showing lung and vertebral involvement. Anti-tuberculosis medication is the first line of treatment in calcaneal TB and surgery is usually reserved for patient with severe, intractable disease. Bangladesh Journal of Infectious Diseases, June 2023;10(1):52-55

Detection of Pulmonary Tuberculosis by Gene Xpert Method among HIV Patient in ART Center, Gadarif State, Sudan

Background: Tuberculosis (TB) is one of the most common and deadly infectious diseases in humans. Due to widespread poverty, inequity, and conflict, suboptimal health services in many countries, and the impact of the HIV/AIDS pandemic, TB/HIV co-infected patients frequently test negative for TB with direct microscopy, posing diagnostic challenges. Traditional HIV-associated TB diagnosis is complex, expensive, time-consuming, and technically demanding because it relies on conventional culture and drug susceptibility testing. The lengthy wait for results has disastrous consequences for patients who go undiagnosed or are diagnosed too late.
 Objective: To detect the accuracy of gene X pert® MTB/RIF as a novel automated molecular tool to diagnose Mycobacterium tuberculosis in HIV patients.
 Methods: A cross sectional study was conducted on 70 HIV patients suspected to have Mycobacterium tuberculosis and who were referred to the ART and Tuberculosis Centers at Gadarif state, from June 2022 to December 2022. Sputum samples were tested by two methods involved the Gene X pert (Cepheid, California, United States of America) and direct smear microscopy (SM) stained with ZN stain for detection of acid fast bacilli. Out of 70 sputum samples analyzed for acid fast bacilli, 50(71%) cases were positive using GeneXpert whereas the DM was positive only for 28 (4o %) of the cases. GeneXpert detected another 22 AFB positive cases. Therefore GeneXpert is more accurate. Sensitivity of the GeneXpert was calculated as 93% and the specificity was 43%. The majority of the patients who contributed to (SM) were found to be co-infected with HIV.
 Conclusion: GeneXpert demonstrated two-time case detection rate compared to the sputum smear microscopy so the direct smear showed poor performance tool for evaluation of TB-HIV co-infection. Also The Gene X pert’s sensitivity was found to be high while the specificity was low. Although this was the case, the GeneXpert as compared to DM could greatly reduce false negatives and the wait on treatment initiation can be significantly shortened, minimising premature death and continued transmission.

Comparative analysis of cartridge-based nucleic acid amplification test (CBNAAT) with conventional methods in the diagnosis of pediatric tuberculosis at a tertiary care center

BackgroundTuberculosis is an important cause of morbidity and mortality among children. Early diagnosis and treatment in children are challenging, more so in resource-limited, tuberculosis-endemic countries. In 2017, the WHO endorsed the use of CBNAAT for tuberculosis diagnosis. We have undertaken this study to evaluate the diagnostic value of CBNAAT in pediatric tuberculosis in comparison to other methods like microscopic detection of acid-fast bacilli and detection of mycobacteria-by-mycobacteria growth indicator tube (MGIT). Material and methodsThis hospital-based, cross-sectional, observational prospective study was conducted in the department of pediatrics, at a tertiary care center. A detailed history, general physical examination, and relevant physical examination were performed systematically and the findings were noted in the proforma. All necessary basic investigations like CBC, ESR, X-Ray, etc., and advanced investigations like MRI, CT, and FNAC were done as per the requirement of the subjects and the results were mentioned in the study proforma. Sensitivity, specificity, positive and negative predictive value, and diagnostic accuracy were calculated for various methods. A comparison between the two methods was done using the Mc Nemar test. p-value ≤0.05 was taken as statistically significant. All statistical analyses were done using Epi info version 7.2.1.0 statistical software. ResultsAmong 102 children suspected to be suffering from tuberculosis, the maximum number of TB cases were found in the age group of 11–16 years (43.2%), there were 58.2% of females, 58.8% belonged to the rural population, fever (78.4%) was the most common presenting symptom and 35.3% had a history of contact. In the present study, CBNAAT and ZN staining had equal sensitivity (60.8%) and specificity (100%) while the yield for MGIT culture was quite low (sensitivity 37.3%, specificity 100%). ConclusionsCBNAAT as a test was found to be useful, especially for early diagnosis and detection of rifampicin resistance in pediatric tuberculosis against MGIT culture. Since MGIT results become available only after 42 days and have a relatively lower yield so they can be utilized only in a selected clinical situation or in patients with high suspicion of tuberculosis where another test is not able to detect the organisms.

Utility of anti-Mycobacterium tuberculosis antibody (ab905) for detection of mycobacterial antigens in formalin-fixed paraffin-embedded tissues from clinically and histologically suggestive extrapulmonary tuberculosis cases

BackgroundThe detection of acid-fast bacilli in extrapulmonary tissue samples is challenging due to its paucibacillary nature. The present study assessed the utility of immunohistochemistry (IHC) using anti-Mycobacterium tuberculosis antibody (ab905) for detecting the presence of mycobacterial antigens in archived formalin-fixed paraffin-embedded (FFPE) tissues. MethodsFFPE tissues [surgical biopsies (n = 32) and post-mortem tissues (n = 8)] from clinically and histologically suggestive extrapulmonary tuberculosis (EPTB) cases at the Korle Bu Teaching Hospital, Accra, Ghana from 2015 to 2020 were stained with IHC (anti-Mycobacterium tuberculosis antibody) and Ziehl-Neelsen (ZN) stain. The staining outcomes of IHC and ZN were compared, and their sensitivity and specificity determined against histopathology as reference standard. ResultsLymph nodes were about 40% (16/40) of the samples analyzed. IHC stained positive in 43.8% (7/16) biopsies and 87.5% (4/5) post-mortem samples ranging from 43.8% (7/16) in lymph nodes to 80% (4/5) in gastrointestinal organs. The overall sensitivity for IHC was 52.50% (95% CI: 36.13%–68.49%) and 0% (95% CI: 0.00%–8.81%) for ZN. Specificity was 72.50% (95% CI: 56.11%–85.40%) and 75% (95% CI: 58.80–87.31%) for IHC and ZN respectively. ConclusionsIHC using anti-Mycobacterium tuberculosis antibody (ab905) can detect mycobacterial antigens in diverse range of paucibacillary extrapulmonary tissue sections. It is potentially a useful tool for the diagnosis of EPTB in FFPE tissues in a routine pathology laboratory.

Pulmonary Granulomas and Mycobacterial Infection: Concordance between the Results of Special Stains Performed on Lung Tissue Sections and Tissue Cultures.

The most common cause of infectious pulmonary granulomas worldwide is Mycobacterium tuberculosis. The diagnosis is based on clinical presentation, histopathologic findings, detection of acid-fast bacilli (AFB) in tissue or sputum using special stains, and/or isolation of mycobacteria in cultures or via PCR-based methods. Different studies have shown that high levels of discrepancy exist between these diagnostic approaches in lung tissue specimens. To assess the degree of concordance between the results of special stains and cultures on lung tissue specimens in the diagnosis of mycobacterial infections. Eighty-seven patients with a diagnosis of granulomas (necrotizing and non-necrotizing) on lung tissue specimens were identified. Cohen's kappa was used to measure the general concordance between the results of the histopathological examination (special stains) and bacteriological tissue cultures. With Kinyoun acid-fast stains, 8/48 (16.7%) cases were positive for AFB. With FITE stains, 10/57 (17.5%) cases were positive for AFB. There was strong agreement between Kinyoun acid-fast and FITE stains (Kappa = 0.806; p-value < 0.001). Tissue cultures were performed on 38/87 cases (43.7%), and 10/38 (26.3%) of the cultures were positive for mycobacteria. There was no concordance between Kinyoun acid-fast stains or FITE stains and tissue cultures results. Our observations represent an initial step in the process of reviewing the two methods used at our institution to diagnose mycobacterial infections on lung tissue specimens and highlight the need of incorporating more advanced diagnostic methods such as PCR to confirm mycobacterial infections and improve patient management. Importantly, species-level identification of mycobacteria is necessary to guide treatment.

Evaluation of phenol ammonium sulfate basic fuchsin and auramine O staining by pot technique for the detection of acid-fast bacilli among patients suspected of pulmonary tuberculosis

Introduction and Aim: Tuberculosis (TB) remains a major public health concern with its high incidence and mortality. In developing countries, smear microscopy continues to be the primary diagnostic tool for the diagnosis of TB. Sputum samples processing under resource-limited settings could be hazardous, which urges safe and efficient smear microscopy techniques. This study aimed to evaluate the efficacy of two different pot methods: Phenol ammonium sulfate (PhAS) auramine O and PhAS basic fuchsin in comparison to the conventional Auramine O method. Materials and Methods: A prospective cross-sectional study was conducted at the Department of Microbiology with 74 sputum samples. All sputum samples were processed using the Auramine O method PhAS auramine O and PhAS basic fuchsin staining methods. Microsoft Office Excel (Microsoft, Redmond, WA, USA) was used to maintain and analyze all the data. Results: A total of 8 (10.7%) samples were positive for AFB and 67 (89.3%) samples were negative, according to routine auramine O method and pot methods. Pot methods showed a 100% sensitivity and specificity compared to the conventional Auramine O method. In terms of sputum smear grading, the direct smear using auramine O showed better results compared to pot methods. Conclusion: Pot methods like PhAS basic fuchsin and PhAS auramine O are efficient to detect AFB in sputum smears by reducing the risk of laboratory-acquired infections.

Evaluation of modified bleach technique for the detection of acid fast bacilli in lymph node aspirate at the University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia

BackgroundTuberculous Lymphadenitis (TBLN) is the commonest form of extrapulmonary tuberculosis. Definitive diagnosis of TBLN is difficult due to the low efficacy of the routine diagnostic techniques as compared to culture. ObjectiveTo determine whether prior bleach concentration can improve the detection of acid-fast bacilli when compared with conventional Ziehl-Neelsen (ZN) staining by using culture as the gold standard. MethodsCross-sectional study was conducted from March 01, 2015 to May 30, 2015. The study subjects were clinically suspected TBLN patients. Fine needle aspiration (FNA) was performed. Two different smears were prepared directly from the first needle pass for cytology and ZN staining. The remaining aspirate was used for the modified bleach concentration. The second needle pass aspirate was transferred into a sterile container containing sterile normal saline. The sediment was inoculated on the Lowenstein Jensen (LJ) medium. Measurement parameters for reliability and validity were used to analyze the results. ResultA total of 93 participants were included in the study. Fifty-six out of the 93 (60.2 %) cases were positive for mycobacterium tuberculosis complex on LJ culture. The detection rates of direct ZN staining and modified bleach method were 20.4 % and 44.1 %, respectively. 73.1 % of the cases showed cytomorphological features consistent with TBLN by cytologic examination. The sensitivities of direct ZN staining and modified bleach method and cytomorphology were 32.0 %, 67.8 %, and 92.8 %, respectively. ConclusionImplementation of bleach concentration increases the detection rate of AFB over the direct ZN method. The bleach method can also be easily performed and provide a safe working environment by reducing infections.

Urine-lipoarabinomannan-might fill the gap in delayed diagnosis of life-threatening tuberculosis

Mycobacterium tuberculosis bacilli is a causative organism for tuberculosis disease. The lungs are the most commonly affected organ with this infection but it can infect any vital organ of the body. The person with signs and symptoms of tuberculosis disease was identified with detection of acid-fast bacilli in the biological specimen and with rapid molecular testing. However, the limiting factor for the early tuberculosis (Tb) diagnosis is the running cost of the diagnostic test. It is the common reason preventing the advancement of diagnostic laboratories in low and middle-income countries. The person and community both suffered from this diagnostic delay. This delay causes significant morbidity and mortality in the patient. An alternate test and an alternate sample that can be easily obtained would be beneficial to prevent these diagnostic delay issues. Lipoarabinomannan (LAM) antigen detection in the urine sample is one such promising diagnostic test. Mycobacterium tuberculosis’s inner layer is made up of glycolipids. LAM is derived from phosphatidylinositol. LAM is a heat-stable amphiphilic cell wall component. It is the precursor of phosphatidyl-inositol-mannosidase and lipomannan. An extra mannose cap is a characteristic feature of a virulent strain of Mycobacterium. LAM has immunogenic and immunomodulatory properties. It is a promising diagnostic test because it is simple to do, the assay can be performed at the patient’s bedside and takes a while to perform. This assay sensitivity varies from 56% to 85% and it has greater than 88% specificity. In HIV seropositive patients, use of LAM assay can reduce 8-week mortality. LAM detection is also a very good assay for the detection of tuberculosis in renal failure and disseminated tuberculosis patients.Lipoarabinomannan detection in the urine is a possible test that can prevent delay in diagnosis. This is a promising test because it's easy to perform, the test can be done with a card test besides the patient's bed. It can be used in HIV seropositive patients and various other forms of extra pulmonary tuberculosis. It can be a very useful test for diagnosing a critically ill patient who is not able to produce a target sample.

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.

Deep-Learning-Aided Detection of Mycobacteria in Pathology Specimens Increases the Sensitivity in Early Diagnosis of Pulmonary Tuberculosis Compared with Bacteriology Tests.

The histopathological diagnosis of mycobacterial infection may be improved by a comprehensive analysis using artificial intelligence. Two autopsy cases of pulmonary tuberculosis, and forty biopsy cases of undetected acid-fast bacilli (AFB) were used to train AI (convolutional neural network), and construct an AI to support AFB detection. Forty-two patients underwent bronchoscopy, and were evaluated using AI-supported pathology to detect AFB. The AI-supported pathology diagnosis was compared with bacteriology diagnosis from bronchial lavage fluid and the final definitive diagnosis of mycobacteriosis. Among the 16 patients with mycobacteriosis, bacteriology was positive in 9 patients (56%). Two patients (13%) were positive for AFB without AI assistance, whereas AI-supported pathology identified eleven positive patients (69%). When limited to tuberculosis, AI-supported pathology had significantly higher sensitivity compared with bacteriology (86% vs. 29%, p = 0.046). Seven patients diagnosed with mycobacteriosis had no consolidation or cavitary shadows in computed tomography; the sensitivity of bacteriology and AI-supported pathology was 29% and 86%, respectively (p = 0.046). The specificity of AI-supported pathology was 100% in this study. AI-supported pathology may be more sensitive than bacteriological tests for detecting AFB in samples collected via bronchoscopy.

Human immunodeficiency virus p24 antigen and antibody, herpes simplex virus-2 IgM and tumor necrosis factor alpha plasma levels in adult female children living in single-parent households: a case-control observational study

Abstract Objective: Human immunodeficiency virus (HIV) p24 antigen and antibody and herpes simplex virus 2 IgM are seromarkers indicating infection with HIV and herpes simplex virus-2 (HSV-2), respectively, whereas tumor necrosis factor α is an inflammatory biomarker that can be triggered by infections. Female children of single parents are faced with many socio-economic challenges that make them vulnerable to sexual influences and prone to sexually transmitted infections. The goal of this work was to determine HIV p24 antigen/antibody, HSV-2 IgM and tumor necrosis factor-α plasma levels in adult female children living in single-parent households. Methods: In this case-control observational study, 100 adult female children living with a single parent (50 living with a single mother and 50 living with a single father; age: 18–22 years) and 100 age-matched women living with both parents were recruited to serve as the test and control groups, respectively. All subjects were negative for acid-fast bacilli, plasmodium, hepatitis C virus, and hepatitis B virus. Human tumor necrosis factor α, HSV-2 IgM, antibody to hepatitis C virus, hepatitis B surface antigen and human immunodeficiency virus p24 antigen and antibody (HIV p24 Ag/Ab) levels were determined by ELISA, while the detection of acid-fast bacilli in sputum and Plasmodium in blood was carried out by optical microscopy. This work was carried out in the Owo/Ose Federal Constituency in Ondo State that shares boundaries with Edo State. The study protocol was approved by the Research and Ethical Committee of the Department of Medical Laboratory Science, Achievers University, Owo, Nigeria (AUO/MLS/2020/127) on August 27, 2020. Results: HIV p24 Ag/Ab was detected in 0 adult female children living with a single mother, 1 (2%) adult female child living with a single father and 1 (1%) adult female child living with both parents. HSV-2 IgM was detected in 9 (18%) adult female children living with a single mother, 13 (26%) adult female children living with a single father, and 5 (10%) adult female children living with both parents. Conclusion: This work shows that adult female children of single parents are vulnerable to sexual influences, and thereby more prone to HSV-2 and possibly HIV, especially adult female children of single fathers.

Hemophagocytic Lymphohistiocytosis and Severe Hypoxemia in a Preterm Infant.

Hemophagocytic Lymphohistiocytosis and Severe Hypoxemia in a Preterm Infant.

Concentrated specimen smear microscopy utilising a polymer membrane sandwich filtration vessel for the detection of acid-fast bacilli in health facilities in Sabah, East Malaysia

A simple, ready-to-use concentrated specimen smear microscopy method employing a nanometer silicon polyvinylidene fluoride (PVDF) polymer membrane sandwich filtration vessel to concentrate acid-fast bacilli (AFB) in samples (SFV-CSSM, Hunan-Tech New Medical System Co. Ltd. China) was compared with direct sputum smear microscopy (DSSM) to determine its performance using culture on modified Ogawa agar as reference. The results for 4114 clinical samples collected from health facilities in Sabah were interpreted with reference to culture results, sample collection-transportation conditions and clinical data including responses to anti-TB drug treatment. The SFV-CSSM showed higher sensitivity than DSSM (79.4% versus 60.5%) and less background interference. Its ability to detect low levels of AFB at an affordable cost makes it an excellent tool for the screening of pauci-bacillary samples as well as for active case finding in TB control programs.

Efficacy of fluorescent microscopy versus modified Fite‐Faraco stain in skin biopsy specimens of leprosy cases – a comparative study

Histopathological examination of skin remains the cornerstone in the diagnosis of leprosy. At a few centers, fluorescent microscopy has been found to be useful in detecting more acid-fast bacilli (AFB) compared to modified Fite-Faraco staining but is sparsely documented. Hence, we studied the sensitivity of fluorescent microscopy and modified Fite-Faraco stain in the detection of Mycobacterium leprae in tissue sections. Patients attending our outpatient department during January 2019 to June 2020 with the clinical features of leprosy were examined, and the diagnosis was confirmed by histopathology after informed consent. Tissue sections were stained by fluorescent stain and modified Fite-Faraco stain. Bacillary index was calculated for each case. Forty patients were recruited after following the inclusion and exclusion criteria. AFB were demonstrated in 20 patients by modified Fite-Faraco stain and in 27 patients with fluorescent stain. The sensitivity of fluorescent staining method (67.5%) was higher than modified Fite-Faraco stain (50%). Bacillary index was increased in 26 out of 40 cases by the fluorescent staining compared to the modified Fite-Faraco staining. Chi-square value was 69.3 and P value was 0.000, indicating a statistically significant correlation. Fluorescent microscope is expensive, and trained people are needed to identify the bacilli. Fluorescent staining is more sensitive than modified Fite-Faraco staining in the detection of AFB in tissue sections. The bacilli detected per field were high with the fluorescent staining compared to the modified Fite-Faraco method.

Tween 80 Improves the Acid-Fast Bacilli Quantification in the Magnetic Nanoparticle-Based Colorimetric Biosensing Assay (NCBA).

Despite its reduced sensitivity, sputum smear microscopy (SSM) remains the main diagnostic test for detecting tuberculosis in many parts of the world. A new diagnostic technique, the magnetic nanoparticle-based colorimetric biosensing assay (NCBA) was optimized by evaluating different concentrations of glycan-functionalized magnetic nanoparticles (GMNP) and Tween 80 to improve the acid-fast bacilli (AFB) count. Comparative analysis was performed on 225 sputum smears: 30 with SSM, 107 with NCBA at different GMNP concentrations, and 88 with NCBA-Tween 80 at various concentrations and incubation times. AFB quantification was performed by adding the total number of AFB in all fields per smear and classified according to standard guidelines (scanty, 1+, 2+ and 3+). Smears by NCBA with low GMNP concentrations (≤1.5 mg/mL) showed higher AFB quantification compared to SSM. Cell enrichment of sputum samples by combining NCBA-GMNP, incubated with Tween 80 (5%) for three minutes, improved capture efficiency and increased AFB detection up to 445% over SSM. NCBA with Tween 80 offers the opportunity to improve TB diagnostics, mainly in paucibacillary cases. As this method provides biosafety with a simple and inexpensive methodology that obtains results in a short time, it might be considered as a point-of-care TB diagnostic method in regions where resources are limited.