Challenges in Diagnosis of Extrapulmonary Tuberculosis

Even in the era of twenty-first century tuberculosis is still considered as a major health burden around the globe particularly around the Indian sub-continent. Though Pulmonary system is the principal site for Mycobacterium but extra-pulmonary involvement is not so common. Extra-pulmonary involvement can be seen in isolation or even with pulmonary involvement also. Immunocompromised individuals like HIV-affected individuals carries a greater risk for disseminated tuberculosis with involvement of multiple extra-pulmonary sites but Immunocompetent persons also can develop extra-pulmonary manifestation. Among all sites, Lymph nodes are the commonest in extra-pulmonary involvement. Though conventional Sputum smear examination and culture sensitivity is still reliable in diagnosis of pulmonary tuberculosis but are less helpful in extra-pulmonary cases due to its paucibacillary nature. Modern molecular methods in background of strong clinical suspicion with or without radiological evidences forms the pathway to confirm the diagnosis. These Diagnostic difficulty makes the delay in response to treatment in these patients

Oral bacteria detection among children with cancer in a tertiary teaching hospital in Kuala Lumpur, Malaysia

@# This study sought to determine the prevalence of pathogenic and non-pathogenic bacteria in the oral cavities of children with cancer. There were 68 paediatric patients with cancer who were included in this study. Oral swab samples from the dorsum of tongues and mouth floors of these patients were subjected to culture, staining, and molecular methods to detect the bacteria. The overall prevalence of gram-positive and gram-negative bacteria was 79.4% (54/68; 95% CI = 68.4 – 87.3) and 25% (17/68; 95% CI = 16.2 – 36.4), respectively. Streptococcus salivarius and Streptococcus parasanguinis were the predominant pathogenic grampositive bacteria, while Neisseria subflava and Neisseria perflava were the most common pathogenic gram-negative bacteria. The results revealed that the number of bacteria isolates recovered in patients receiving cancer treatment was higher (55.9%) than those who had not received treatment (16.2%). Therefore, more isolated pathogenic bacteria were observed post-therapy (54.4%). Pathogenic organisms can have significant implications on patient health. Awareness of the types of bacteria inhabiting the oral cavity is essential to predict and prevent dental problems, and their associated systemic complications. Findings on the diversity of oral microflora can also provide a better understanding of the aetiology of oral diseases in paediatric patients receiving cancer treatment.

Molecular methods for detection of pathogenic viruses of respiratory tract - A review

Human respiratory system is harbored by a vast array of transient and normal microbial flora. A number of pathogenic viruses were diagnosed from samples in different occasions from mild to severe infections of respiratory tract. Molecular methods were developed for detection of these viruses during last two decades. Nucleic acid amplification methods were introduced for rapid and accurate diagnosis of pathogenic viruses. Multiplex detection systems were employed to identify a panel of pathogenic viruses, which requires specialized kits and instruments in some cases. This review summarizes different types of molecular approaches which were developed with time and applied for the detection of pathogenic viruses associated with infections of the respiratory system.

The First Case of Diarrhoea in Tibetan Sheep, Ovis aries, Caused by Balantidium coli in the Qinghai Tibetan Plateau Area, China

This study was carried out to determine the pathogen-causing diarrhoea in sheep Ovis aries in the Qinghai Tibetan Plateau Area, China. A trophozoite was identified as species of ciliate alveolates infecting the sheep based on morphological characteristics examined by microscope. It was mostly spherical, colourless and transparent, with many vesicles. Macronucleus and contractile vacuoles could not be distinguished. Size of the trophozoite was 80–180×70–150 μm and its surface was covered with cilia. Molecular analysis based on sequences of 18S rRNA and ITS genes confirmed the ciliate species as Balantidium coli. According to the literature, there have been many epidemiological investigations of B. coli infection in pigs, monkeys and humans. To our knowledge, this was the first report of B. coli infections in sheep in the Qinghai Tibetan Plateau Area of China, or eleswhere around the world. Importantly, the sheep case was rare but raised our concern that B. coli may spread across species and expand its host range.

Molecular methods for detection of pathogenic viruses of respiratory tract-A review

Human respiratory system is harbored by a vast array of transient and normal microbial flora. Anumber of pathogenic viruses were diagnosed from samples in different occasions from mild tosevere infections of respiratory tract. Molecular methods were developed for detection of theseviruses during last two decades. Nucleic acid amplification methods were introduced for rapidand accurate diagnosis of pathogenic viruses. Multiplex detection systems were employed toidentify a panel of pathogenic viruses, which requires specialized kits and instruments in somecases. This review summarizes different types of molecular approaches which were developedwith time and applied for the detection of pathogenic viruses associated with infections of therespiratory system.

A primary assessment of the endophytic bacterial community in a xerophilous moss (Grimmia montana) using molecular method and cultivated isolates

Investigating the endophytic bacterial community in special moss species is fundamental to understanding the microbial-plant interactions and discovering the bacteria with stresses tolerance. Thus, the community structure of endophytic bacteria in the xerophilous moss Grimmia montana were estimated using a 16S rDNA library and traditional cultivation methods. In total, 212 sequences derived from the 16S rDNA library were used to assess the bacterial diversity. Sequence alignment showed that the endophytes were assigned to 54 genera in 4 phyla (Proteobacteria, Firmicutes, Actinobacteria and Cytophaga/Flexibacter/Bacteroids). Of them, the dominant phyla were Proteobacteria (45.9%) and Firmicutes (27.6%), the most abundant genera included Acinetobacter, Aeromonas, Enterobacter, Leclercia, Microvirga, Pseudomonas, Rhizobium, Planococcus, Paenisporosarcina and Planomicrobium. In addition, a total of 14 species belonging to 8 genera in 3 phyla (Proteobacteria, Firmicutes, Actinobacteria) were isolated, Curtobacterium, Massilia, Pseudomonas and Sphingomonas were the dominant genera. Although some of the genera isolated were inconsistent with those detected by molecular method, both of two methods proved that many different endophytic bacteria coexist in G. montana. According to the potential functional analyses of these bacteria, some species are known to have possible beneficial effects on hosts, but whether this is the case in G. montana needs to be confirmed.

Performance Assessment of Advansure(TM) MDR-TB Genoblot Assay Kit for Anti-tuberculosis Drug Susceptibility Test

BACKGROUND: Because of the long time required for conventional drug susceptibility test (DST) for rifampin and isoniazid, development of rapid DSTs is necessary. Recently, the AdvanSure(TM) MDR-TB GenoBlot Assay kit (LG Life Science, Korea), using reverse hybridization line blot assay, was developed. We compared this kit with Genotype(R) MTBDRplus (HAIN Lifescience, Germany) and conventional DST. METHODS: Of the DNAs preserved after performing DST by using Genotype(R), we selected 144 samples having conventional DST results. The experiments with both the kits were performed according to the manufacturers' instructions. For the samples for which discrepant results were obtained, sequencing was performed if the DNA was available. Conventional DST was performed at the Korean Institute of Tuberculosis by using the absolute concentration method. RESULTS: For rifampin, the findings obtained using both the kits were the same with concordance rates of 98.6% (142/144) compared to conventional DST. Of the 2 discrepant findings, one was very major error and the other was major error. For isoniazid, compared to conventional DST, concordance rates of AdvanSure(TM) and Genotype(R) were 95.8%(138/144) and 95.1%(137/144) respectively. Of the 6 discrepant findings between conventional method and Advansure(TM), 5 were very major error and one was major error. All the 7 discrepant findings between conventional method and Genotype(R) were very major error. CONCLUSIONS: The findings obtained using AdvanSure(TM) showed high concordance with those obtained using Genotype(R) and conventional DST. This kit has a higher rate of detection of isoniazid resistance because it includes probes for an additional target (ahpC).