Research Article Open Access Riyaz et al., J Med Microb Diagn 2019, 8:1 DOI: 10.4172/2161-0703.1000294 Research Article Open Access Journal of Medical Microbiology & Diagnosis Volume 8 • Issue 1 • 1000294 J Med Microb Diagn, an open access journal ISSN: 2161-0703 J o u r n a l o f M e d i c a l M i c r o b i o l o g y & D i a g n o s i s ISSN: 2161-0703 *Corresponding author: Nazish Fatima, Department of Microbiology, Jawahar Lal Nehru Medical College, Aligarh Muslim University, Uttar Pradesh, India, Tel: +919412174543; E-mail: nazsham28@gmail.com Received December 28, 2018; Accepted January 27, 2019; Published February 16, 2019 Citation: Riyaz S, Fatima N, Khan HM, Shameem M (2019) Comparison of Various Laboratory Detection Methods for Diagnosing Pulmonary Aspergillosis. J Med Microb Diagn 8: 294. doi:10.4172/2161-0703.1000294 Copyright: © 2019 Riyaz S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Aspergillus is a mould which may lead to a variety of infectious, allergic diseases depending on the immune status or pulmonary structure of the host. Invasive pulmonary aspergillosis occurs primarily in patients with severe immunodeficiency. Early and rapid diagnosis of systemic fungal infections remain limited, despite intensive efforts by many investigators. Few clinical guidelines have been previously proposed for either diagnosis or management of Pulmonary Aspergillosis. The current study was undertaken for comparing various detection methods like conventional direct microscopy or histopathology, culture methods, immunological (GM ELISA) and molecular methods (PCR). To identify carcinoma cell types and to categorize Aspergillosis types histopathology was done. For direct fungal examination, culture, Aspergillus polymerase chain reaction (PCR), and galactomannan (GM) detection, bronchoalveolar lavage (BAL) fluids were collected. In this study microscopy was found to have low sensitivity of (70.5%) while culture had highest specificity (97.6%). GM assay showed a sensitivity of 100% and a specificity of 86.4% whereas PCR has an overall sensitivity of 100% and a specificity of 81.3%. Thus, we suggest that both BAL PCR and GM ELISA may be beneficial for use in early diagnosis of Aspergillosis, especially those patients who do not demonstrate radiological signs. Comparison of Various Laboratory Detection Methods for Diagnosing Pulmonary Aspergillosis Sadaf Riyaz 1 , Nazish Fatima 1* , Harris M Khan 1 and Shameem M 2 1 Department of Microbiology, Jawahar Lal Nehru Medical College, Aligarh Muslim University, Uttar Pradesh, India 2 Department of TB and Respiratory Diseases, Jawahar Lal Nehru Medical College, Aligarh Muslim University, Uttar Pradesh, India Keywords: Pulmonary Aspergillosis; Diagnosis; PCR; ELISA Introduction IPA was first described in 1953 [1]. Due to widespread use of chemotherapy and immunosuppressive agents, its incidence has increased over the past two decades [2]. Despite intensive efforts by many investigators, early and rapid diagnosis of systemic fungal infections remain limited. Aspergillus spp. when isolated from respiratory samples does not confirm it as the etiologic pathogen because airway colonization by Aspergillus spp. is a common feature in several chronic lung diseases. Only if Aspergillus spp. is repeatedly isolated and anti- Aspergillus antibodies and/or Aspergillus antigens in sera are detected, it points towards the etiologic agent of disease [3]. e only available techniques in most centers to diagnose IA are conventional direct microscopy, histopathology, and culture methods, due to the non-availability of galactomannan or beta-glucan tests assay in developing countries [4]. Bronchoscopy with Bronchoalveolar Lavage (BAL) is generally helpful in diagnosis of IPA especially in patients with diffuse lung involvement [5]. Generally, clinical and radiologic signs are insensitive and non-specific, and the sensitivity of fungal culture is low [5]. Histopathology plays a good diagnostic role in IPA, but the invasive nature of tissue biopsy collection discourages its use in thrombocytopenic patients. us, rapid and more sensitive diagnostic strategies for fungal infections including detection of antigen or DNA are being evaluated [6,7]. Galactomannan (GM), double sandwich ELISA (Platelia, Bio-rad) which incorporates the β1-5 Galactofuranose specific EBA2 monoclonal antibody as both the acceptor and the detector for Galactomannan, a polysaccharide cell-wall component that is released by Aspergillus during growth, has the most promise among the many tests that have been developed for detection [7]. In the Revised Criteria of EORTC/ MSG for probable invasive Aspergillosis, the Galactomannan antigen detection has also been included. Although, the detection of nucleic acid is not included in the criteria, as presently, there are no validated or standardized methods [8]. While GM cannot identify infecting Aspergillus species, PCR could be tailored to the species level and may infer general antifungal susceptibility pattern [5]. Materials and Methods is study was conducted on patients admitted in the wards or attending the outpatient department of T.B. and Respiratory Diseases, Jawaharlal Nehru Medical College and Hospital, AMU, Aligarh, for a period of one and half year. Study group comprised of patients suspected of lung carcinoma and chronic lung diseases like interstitial lung disease, chronic obstructive pulmonary disease etc. Bronchoscopy was performed for confirmation of clinical diagnosis in around 60 patients. Age and sex matched healthy controls were also included in the study with no evidence of any chronic lung disease. A detailed clinical history was recorded for each patient especially regarding duration of illness, occupation (especially any exposure to grains), history of smoking/gutka chewing, dietary history and others like: Signs and symptoms, broad spectrum antibiotics and corticosteroid therapy. In a clean sterile vial, Bronchoalveolar lavage (BAL) was collected, by fiberoptic bronchoscopy. Each BAL specimen was divided in four parts for direct microscopy, culture, galactomannan and PCR. Each specimen of bronchoalveolar lavage was subjected to various laboratory procedures for isolation and identification of Aspergillus spp.