Establishment of reference CD4+ T cell values for adult Indian population

<p>Abstract</p> <p>Background</p> <p>CD4+ T lymphocyte counts are the most important indicator of disease progression and success of antiretroviral treatment in HIV infection in resource limited settings. The nationwide reference range of CD4+ T lymphocytes was not available in India. This study was conducted to determine reference values of absolute CD4+ T cell counts and percentages for adult Indian population.</p> <p>Methods</p> <p>A multicentric study was conducted involving eight sites across the country. A total of 1206 (approximately 150 per/centre) healthy participants were enrolled in the study. The ratio of male (N = 645) to female (N = 561) of 1.14:1. The healthy status of the participants was assessed by a pre-decided questionnaire. At all centers the CD4+ T cell count, percentages and absolute CD3+ T cell count and percentages were estimated using a single platform strategy and lyse no wash technique. The data was analyzed using the Statistical Package for the Social Scientist (SPSS), version 15) and Prism software version 5.</p> <p>Results</p> <p>The absolute CD4+ T cell counts and percentages in female participants were significantly higher than the values obtained in male participants indicating the true difference in the CD4+ T cell subsets. The reference range for absolute CD4 count for Indian male population was 381-1565 cells/μL and for female population was 447-1846 cells/μL. The reference range for CD4% was 25-49% for male and 27-54% for female population. The reference values for CD3 counts were 776-2785 cells/μL for Indian male population and 826-2997 cells/μL for female population.</p> <p>Conclusion</p> <p>The study used stringent procedures for controlling the technical variation in the CD4 counts across the sites and thus could establish the robust national reference ranges for CD4 counts and percentages. These ranges will be helpful in staging the disease progression and monitoring antiretroviral therapy in HIV infection in India.</p

Paragonimus

Paragonimiasis or endemic hemoptysis is an important disease\ud of man and other mammals caused by several trematode species belonging to the genus Paragonimus. Typically,\ud paragonimiasis is a disease of the lungs and pleural cavity\ud but extra-pulmonary paragonimiasis is also an important\ud clinical manifestation. Lung-flukes utilize a mollusk as a first intermediate host and a freshwater crab or crayfish as a second intermediate host. Consumption of raw, undercooked or pickled freshwater crab or crayfish introduces lung-flukes into the human body. Similarly, raw or undercooked meat of paratenic hosts such as boar, bear, wild pig or rat, in the muscles of which juvenile worms can survive for years, is also an important source of human infection.\ud \ud Paragonimiasis is a neglected disease that has received\ud relatively little attention from public health authorities. It has been estimated that around 293 million people are at risk and several million are infected worldwide. However, this may be an underestimate as there are still many places where the disease burden has yet to be assessed. In recent times there has been increased recognition of the public health importance of paragonimiasis and other foodborne trematodiases. In the case of paragonimiasis, this resurgence of interest is partly due to the common diagnostic confusion of paragonimiasis with tuberculosis. Symptoms of the former closely mimic those of tuberculosis, frequently leading to inappropriate treatment being administered especially in areas where both tuberculosis and paragonimiasis co-occur and are overlapping public health issues? Failure of patients to respond to treatment for tuberculosis may lead to inflated estimates of the prevalence of multi-drug resistant tuberculosis and have other far-reaching health implications. Interest in Paragonimus species outside endemic areas is also increasing\ud because of the risk of infection through consumption of\ud crustaceans traded far from their point of origin in today's\ud globalized food supply. \ud \ud Traditionally, morphological criteria were used for identification and taxonomy of lung-flukes. However, the explosion of molecular techniques in recent years has led to the development of better diagnostic tools and a better understanding of lung-fluke taxonomy. Nevertheless, there is still confusion regarding the taxonomic status of a number of species. Molecular characterization of lung-flukes will improve our understanding of evolution and epidemiology of paragonimiasis and our capacity to control this foodborne disease

Presence of three distinct genotypes within the Paragonimus westermani complex in northeastern India

The name Paragonimus westermani (Kerbert, 1878) is commonly applied to members of a species complex that includes the well-known Asian lung fluke of medical and veterinary importance. Unambiguous molecular and morphological evidence showing the presence of a member of the complex in India has recently been published. In the present study we report the occurrence of 2 more members of the P. westermani complex in northeastern (NE) India. Surveys of the freshwater crabs Maydelliatelphusa lugubris in NE India revealed 2 morphologically distinct types of lung fluke metacercariae. Phylogenetic analyses, using DNA sequences from ITS2, 28S and cox1 gene regions indicate that these lung metacercariae belong to P. westermani complex. Type 1 metacercariae have a more basal position within the complex whereas type 2 metacercariae are closely related to the relatively derived forms of P. westermani from NE Asia (Japan, Korea, China) and Vietnam. A third type of metacercaria (type 3), detected in another crab host, Sartoriana spinigera in Assam, was phylogenetically close to P. siamensis, also a member of the P. westermani group. Molecular evidence has demonstrated the existence of 3 genotypes of lung flukes within the Paragonimus westermani complex in NE India. Two of these were previously unknown

Morphological and molecular characterization of Paragonimus westermani in northeastern India

Evidence for the presence of lung flukes of the Paragonimus westermani in India remains scant. In particular, evidence based on morphology of adult worms is lacking. Metacercariae of the genus Paragonimus, recovered from crabs in two regions of northeastern India, were raised to adulthood in laboratory rats. Morphologically, these worms appear to be P. westermani. DNA sequences from the second internal transcribed spacer (ITS2) and a portion of the ribosomal large subunit gene (28S) of the nuclear ribosomal RNA gene repeat, as well as fragments of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes, all supported this identification. Molecular phylogenetic methods were used for studying the relatedness of these Indian flukes with counterparts from southeast and far-east Asia. Molecular data showed that Indian representatives of the P. westermani complex represent a distinct lineage. It is unclear whether the Indian form can cause disease in humans as some members of the complex do elsewhere

Pleuropulmonary paragonimiasis due to Paragonimus heterotremus: molecular diagnosis, prevalence of infection and clinicoradiological features in an endemic area of northeastern India

In the northeastern region of India, paragonimiasis is emerging as an important public health problem. However, until now the identity of the species causing human infection has been uncertain and there has been little information on the prevalence and clinicoradiological features of infection in the community. Parasitological and immunological surveys revealed that paragonimiasis was hyperendemic in parts of Arunachal Pradesh. Egg positivity in the sputum was 20.9% and 4.1% in children (age ≤15 years) and adults (age >15 years), respectively. Antibody positivity against excretory–secretory antigen of the adult worm in children and adults was 51.7% and 18.7%, respectively. Chronic cough (97.2%) and haemoptysis (83.3%) were common respiratory symptoms among egg-positive cases. Chest radiography (n = 68) images from egg-positive cases showed that air space consolidation (75%), cavitary lesions (14.7%) and mediastinal adenopathy (11.8%) were very frequent. Less frequent findings were nodular lesions, bronchiectasis, mediastinal adenopathy, pleural thickening and pleural effusion. DNA extracted from eggs from the sputum of patients from Arunachal Pradesh was sequenced. Analyses of the second internal transcribed spacer (ITS2) of nuclear rDNA revealed that the species responsible is Paragonimus heterotremus

Paragonimiasis in tuberculosis patients in Nagaland, India

Background: One of the infections that mimic tuberculosis (TB) is paragonimiasis (PRG), a foodborne parasitic disease caused by lung flukes of the genus Paragonimus. In the northeastern states of India, TB and PRG are endemic; however, PRG is rarely included in the differential diagnosis of TB. Objective: To address limited evidence on the dual burden of TB and PRG in northeastern India, we aimed to document the prevalence of PRG among TB patients using sputum smear, stool examination for children <15 years and ELISA. Design: A cross-sectional study of patients receiving TB treatment in the Médecins Sans Frontières (MSF)-supported TB programme in Mon district, in collaboration with the Regional Medical Research Centre (RMRC), Dibrugarh, Assam, between November 2012 and December 2013. Results: Of 96 patients screened between November 2012 and December 2013, three (3%) had pulmonary PRG and were successfully treated with praziquantel. Conclusions: PRG should be considered in the TB diagnostic algorithms in PRG–TB dual burden areas. In case of TB–PRG co-infection, it is preferable to treat PRG first followed by anti-TB treatment a few days later

Genetic Diversity of <i>Mycobacterium tuberculosis</i> Isolates from Assam, India: Dominance of Beijing Family and Discovery of Two New Clades Related to CAS1_Delhi and EAI Family Based on Spoligotyping and MIRU-VNTR Typing

<div><p>Tuberculosis (TB) is one of the major public health concerns in Assam, a remote state located in the northeastern (NE) region of India. The present study was undertaken to explore the circulating genotypes of <i>Mycobacterium tuberculosis</i> complex (MTBC) in this region. A total of 189 MTBC strains were collected from smear positive pulmonary tuberculosis cases from different designated microscopy centres (DMC) from various localities of Assam. All MTBC isolates were cultured on Lowenstein-Jensen (LJ) media and subsequently genotyped using spoligotyping and 24-loci mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing. Spoligotyping of MTBC isolates revealed 89 distinct spoligo patterns. The most dominant MTBC strain belonged to Beijing lineage and was represented by 35.45% (n = 67) of total isolates, followed by MTBC strains belonging to Central Asian-Delhi (CAS/Delhi) lineage and East African Indian (EAI5) lineage. In addition, in the present study 43 unknown spoligo patterns were detected. The discriminatory power of spoligotyping was found to be 0.8637 based on Hunter Gaston Discriminatory Index (HGDI). On the other hand, 24-loci MIRU-VNTR typing revealed that out of total 189 MTBC isolates from Assam 185 (97.9%) isolates had unique MIRU-VNTR profiles and 4 isolates grouped into 2 clusters. Phylogenetic analysis of 67 Beijing isolates based on 24-loci MIRU-VNTR typing revealed that Beijing isolates from Assam represent two major groups, each comprising of several subgroups. Neighbour-Joining (NJ) phylogenetic tree analysis based on combined spoligotyping and 24-loci MIRU-VNTR data of 78 Non-Beijing isolates was carried out for strain lineage identification as implemented by MIRU-VNTR<i>plus</i> database. The important lineages of MTBC identified were CAS/CAS1_Delhi (41.02%, n = 78) and East-African-Indian (EAI, 33.33%). Interestingly, phylogenetic analysis of orphan (23.28%) MTBC spoligotypes revealed that majority of these orphan isolates from Assam represent two new sub-clades Assam/EAI and Assam/CAS. The prevalence of multidrug resistance (MDR) in Beijing and Non-Beijing strains was found to be 10.44% and 9.01% respectively. In conclusion, the present study has shown the predominance of Beijing isolates in Assam which is a matter of great concern because Beijing strains are considered to be ecologically more fit enabling wider dissemination of <i>M</i>. <i>tuberculosis</i>. Other interesting finding of the present study is the discovery of two new clades of MTBC isolates circulating in Assam. More elaborate longitudinal studies are required to be undertaken in this region to understand the transmission dynamics of MTBC.</p></div