An algorithm for detection of tuberculosis bacilli in Ziehl-Neelsen sputum smear images

This work proposes an algorithm oriented to the detection of tuberculosis bacilli in digital images of sputum samples, inked with the Ziehl Neelsen method and prepared with the direct, pellet and diluted pellet methods. The algorithm aims at automating the optical analysis of bacilli count and the calculation of the concentration level. Several algorithms have been proposed in the literature with the same objective, however, in no case is the performance in sensitivity and specificity evaluated for the 3 preparation methods. The proposed algorithm improves the contrast of the colors of interest, then thresholds the image and segments by labeling the objects of interest (bacilli). Each object then has its geometrical descriptors and photometric descriptors. With all this, a characteristic vector is formed, which are used in the training and classification process of an SVM. For the training 225 images obtained by the 3 preparation methods were used. The proposed algorithm reached, for the direct method, a sensitivity level of 93.67% and a specificity level of 89.23%. In the case of the Pellet method, a sensitivity of 92.13% and a specificity of 82.58% was obtained, while for diluted Pellet the sensitivity was 92.81% and the specificity 83.61%

Examination of Acid-Fast Bacilli in Sputum Using Modified Light Microscope with Homemade Light Emitting Diode Additional Attachment

Typical clinical symptoms and chest X-ray is a marker of Tuberculosis (TB) sufferers. However, the diagnosis of TB in adults should be supported by microscopic examination. Currently, Bacilli microscopic examination of acid-fast bacilli (AFB) in sputum by Ziehl-Neelsen (ZN) coloring is the most widely used. However, for reasons of convenience, especially for laboratories with a considerable amount of smear samples, and due to higher sensitivity compared with ZN staining, the World Health Organization (WHO) has recommended the use of auramine-O-staining (fluorochrome  staining), which is visualized by light emitting diode (LED) fluorescence microscopy. The aim of this study was to evaluate the performance of modified light microscope with homemade LED additional attachment for examination of AFB in sputum using auramine-O-staining method. We compared the sensitivity and specificity of 2 kinds of AFB in sputum methods: ZN and fluorochrome, using culture on Lowenstein-Jensen media as the gold standard. The results showed auramine-O-staining gives more proportion of positive findings (81%) compared to the ZN method (70%). These results demonstrated that the sensitivity of auramine-O-staining was higher than ZN, however it gives more potential false positive results than ZN. The sensitivity of auramine-O-staining in detecting AFB in sputum was 100% while the specificity was 88%

Automated methods for tuberculosis detection/diagnosis : a literature review

Funding: Welcome Trust Institutional Strategic Support fund of the University of St Andrews, grant code 204821/Z/16/Z.Tuberculosis (TB) is one of the leading infectious causes of death worldwide. The effective management and public health control of this disease depends on early detection and careful treatment monitoring. For many years, the microscopy-based analysis of sputum smears has been the most common method to detect and quantify Mycobacterium tuberculosis (Mtb) bacteria. Nonetheless, this form of analysis is a challenging procedure since sputum examination can only be reliably performed by trained personnel with rigorous quality control systems in place. Additionally, it is affected by subjective judgement. Furthermore, although fluorescence-based sample staining methods have made the procedure easier in recent years, the microscopic examination of sputum is a time-consuming operation. Over the past two decades, attempts have been made to automate this practice. Most approaches have focused on establishing an automated method of diagnosis, while others have centred on measuring the bacterial load or detecting and localising Mtb cells for further research on the phenotypic characteristics of their morphology. The literature has incorporated machine learning (ML) and computer vision approaches as part of the methodology to achieve these goals. In this review, we first gathered publicly available TB sputum smear microscopy image sets and analysed the disparities in these datasets. Thereafter, we analysed the most common evaluation metrics used to assess the efficacy of each method in its particular field. Finally, we generated comprehensive summaries of prior work on ML and deep learning (DL) methods for automated TB detection, including a review of their limitations.Publisher PDFPeer reviewe

Examination of Acid-Fast Bacilli in Sputum Using Modified Light Microscope with Homemade Light Emitting Diode Additional Attachment

Typical clinical symptoms and chest X-ray is a marker of Tuberculosis (TB) sufferers. However, the diagnosis of TB in adults should be supported by microscopic examination. Currently, Bacilli microscopic examination of acid-fast bacilli (AFB) in sputum by Ziehl-Neelsen (ZN) coloring is the most widely used. However, for reasons of convenience, especially for laboratories with a considerable amount of smear samples, and due to higher sensitivity compared with ZN staining, the World Health Organization (WHO) has recommended the use of auramine-O-staining (fluorochrome  staining), which is visualized by light emitting diode (LED) fluorescence microscopy. The aim of this study was to evaluate the performance of modified light microscope with homemade LED additional attachment for examination of AFB in sputum using auramine-O-staining method. We compared the sensitivity and specificity of 2 kinds of AFB in sputum methods: ZN and fluorochrome, using culture on Lowenstein-Jensen media as the gold standard. The results showed auramine-O-staining gives more proportion of positive findings (81%) compared to the ZN method (70%). These results demonstrated that the sensitivity of auramine-O-staining was higher than ZN, however it gives more potential false positive results than ZN. The sensitivity of auramine-O-staining in detecting AFB in sputum was 100% while the specificity was 88%

Missed Opportunities for the Diagnosis of Pulmonary Tuberculosis in Ashanti and Western Regions of Ghana; Polymerase Chain Reaction, A Better Tool

The diagnosis of Pulmonary Tuberculosis (PTB) has mainly relied on sputum microscopy and culture. The use of molecular techniques such as the Polymerase Chain Reaction (PCR) and its overwhelming advantages compared with conventional diagnostic methodologies cannot be overemphasized. The main aim of this work was to compare the diagnostic sensitivities of microscopy, culture and PCR. Suspected PTB sputum samples were prospectively collected from six hospitals in the Ashanti and Western regions of Ghana. Microscopy was carried out on all samples at the field sites. Apart from culture and PCR, repeat sputum microscopy was carried out in the laboratories of the Kumasi Centre for Collaborative Research (KCCR). Out of the total 425 cases recruited for the study, 123 (29.0%) were smear positive on site in contrast to 275 (64.7%) positivity rate at KCCR. Regarding culture, 254 (59.9%) samples were culture positive whilst PCR technique using INS 1&2 and PR 8&9 primers were positive in 59.9 % and 56% of cases respectively. The proportion of missed positive cases of microscopy were 131 (51%) compared with culture and 1.1% missed cases when compared with PCR. The sensitivity and specificity of microscopy were 123/254 (48.4%) and 154/302 (50.99%). The study confirms the superiority of PCR in the diagnosis of PTB and indicates that a substantial proportion of PTB cases are missed when microscopy alone is used. In areas where the incidence of PTB is high and at referral hospitals, PCR can be done to augment the diagnosis of TB.Keywords: Pulmonary Tuberculosis, Sputum Microscopy, Culture, Polymerase Chain Reactio

Analysis of Tuberculosis using Smear Image

An automatic method for the detection of Tuberculosis (TB) bacilli from microscopic sputum smear images is presented in this paper. According to WHO, TB is the ninth leading cause of death all over the world. There are various techniques to diagnose TB, of which conventional microscopic sputum smear examination is considered. However, the mentioned method of diagnosis is time intensive and error prone, even in experienced hands. The proposed method performs detection of TB, by image binarization and subsequent classification of detected regions using a convolutional neural network. We have evaluated our gist algorithm using a dataset of sputum smear microscopic images with different backgrounds (high density and low-density images). Experimental results show that the proposed algorithm achieves for the TB detection. The proposed method automatically detects whether the sputum smear images is infected with TB or not. This method will aid clinicians to predict the disease accurately in a short span of time, thereby helping in improving the clinical outcome

Diversity of Mycobacterium tuberculosis in the Middle Fly District of Western Province, Papua New Guinea: microbead-based spoligotyping using DNA from Ziehl-Neelsen-stained microscopy preparations

Tuberculosis remains the world’s leading cause of death from an infectious agent, and is a serious health problem in Papua New Guinea (PNG) with an estimated 36,000 new cases each year. This study describes the genetic diversity of Mycobacterium tuberculosis among tuberculosis patients in the Balimo/Bamu region in the Middle Fly District of Western Province in PNG, and investigates rifampicin resistance-associated mutations. Archived Ziehl-Neelsen-stained sputum smears were used to conduct microbead-based spoligotyping and assess genotypic resistance. Among the 162 samples included, 80 (49.4%) generated spoligotyping patterns (n = 23), belonging predominantly to the L2 Lineage (44%) and the L4 Lineage (30%). This is consistent with what has been found in other PNG regions geographically distant from Middle Fly District of Western Province, but is different from neighbouring South-East Asian countries. Rifampicin resistance was identified in 7.8% of the successfully sequenced samples, with all resistant samples belonging to the L2/Beijing Lineage. A high prevalence of mixed L2/L4 profiles was suggestive of polyclonal infection in the region, although this would need to be confirmed. The method described here could be a game-changer in resource-limited countries where large numbers of archived smear slides could be used for retrospective (and prospective) studies of M. tuberculosis genetic epidemiology