Molecular detection of antimalarial resistance in Plasmodium vivax isolates from a tertiary care setting in Puducherry.

PURPOSE: The study was aimed at detecting the mutation patterns in the drug targets in Plasmodium vivax that confer resistance to the common antimalarial agents used in India. METHODS: A total of 27 Plasmodium vivax isolates collected from whole blood samples over a three year period were subjected to PCR amplification followed by sequencing of the genes pvmdr1, pvdhfr, pvdhps and pvk12, which serve as the molecular targets to detect resistance to chloroquine, pyrimethamine, sulfadoxine and artemisinin respectively. RESULTS: The study found T958 M F1076L double mutants of pvmdr1 in 52 %(14/27) isolates, S58R S117 N double mutants of pvdhfr in 67 % (18/27) isolates, A383G A553G double mutant pvdhps in 59 % (16/27) isolates and wild type of pvk12 gene in all the isolates. CONCLUSIONS: There was a rise in the proportion of double mutants of pvmdr1 and pvdhfr over time. Those cases with double mutant pvmdr1 gene in their isolates were found to have a prolonged hospital stay compared to those without, indicating reduced clinical response to chloroquine.

Performance of microscopy compared to conventional PCR in identification of soil-transmitted helminth infections among antenatal women in a low-prevalence setting.

PURPOSE: Traditional microscopy-based methods may provide inaccurate estimates of Soil transmitted helminth (STH) infections in mild intensity of infection. Therefore, we aimed to determine the prevalence of STH infections using molecular diagnostic methods and compare the diagnostic performance of microscopy with polymerase chain reaction (PCR) in stool samples collected from pregnant women in primary care settings in Puducherry, India. METHODOLOGY: A singleplex PCR assay was developed to detect three species of STHs, namely Ascaris lumbricoides, Necator americanus, and Ancylostoma duodenale, by targeting the internal transcribed spacer regions (ITS1 and ITS2) of 5.8S rRNA. The PCR generated 420, 662, and 515 base pairs of DNA for the respective organisms. In addition to singleplex PCR, wet and concentration microscopy techniques were used. The results were expressed as percentages with 95% confidence intervals, and the diagnostic performance of microscopy was compared with PCR in terms of sensitivity, specificity, and positive, negative predictive values and kappa statistics. RESULTS: Among the 650 pregnant women included, 48.8% were aged 25 years or less, 59% were primigravida, and half were from rural areas. The overall prevalence of any STH infection was higher in PCR compared to microscopy (8.9% vs. 7.2%). The prevalence of Ascaris lumbricoides was higher by microscopy (5.4% vs 2.6%), while the prevalence of Necator americanus was higher by PCR (6.3%) than by microscopy (1.8%). No species of Ancylostoma duodenale was detected. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of microscopy for detecting any STH infection was 22.4%, 94.3%, 27.7%, and 92.5%, respectively. The agreement between microscopy and PCR for the identification is as follows: for any STH infection, k â€‹= â€‹0.12, Ascaris k â€‹= â€‹0.16, and Necator k â€‹= â€‹0.20, respectively. CONCLUSION: The prevalence of any STH infection identified by PCR was higher than microscopy, and the agreement between the two methods was poor.

Prevalence of malaria: A 7-year trend analysis from a tertiary care center, Puducherry.

Background: Battle against malaria has been going on since time immemorial. Understanding the true burden of disease and the determinants of its transmission are important for implementing adequate control measures. This study intends to explore the local epidemiology and burden of malaria in Puducherry, a coastal Union territory located in the Southern part of India over a period of 7 years. Methodology: A retrospective record-based study was conducted from 2015 to 2021, where details from all samples that tested positive for malaria by peripheral blood examination or rapid card test, from suspected cases were collected and analyzed. Results: The overall prevalence of malaria over the 7 years was 1.7% (257/14,888). Majority of the patients were male (75.88%) and the major age group affected was from 21 to 40 years (56.03%). The disease was maximum seen during the monsoon season followed by the post-monsoon season. Vivax malaria predominated irrespective of the gender, seasonal change, and different age groups except in children <10 years was both falciparum and vivax malaria were seen in equivalence. The major species to cause infection among infants were Plasmodium falciparum (3/4). Discussion and Conclusion: This study shows a declining trend of malaria transmission over the years. There is no change in the predominant species affected or seasonal trends over the years. The possibility of underestimation of cases due to various factors cannot be ignored.

Development of a new multiplex PCR to detect fecal coccidian parasite.

BACKGROUND: Cryptosporidium spp., Cystoisospora belli and Cyclospora cayetanensis are common intestinal coccidian parasites causing gastroenteritis. The clinical presentation caused by each parasite is indistinguishable from each other. Uniplex polymerase chain reaction (PCR) for these three groups of intestinal coccidian parasites was developed by us in our laboratory. Thereafter, we planned to develop a single-run multiplex polymerase chain reaction (mPCR) assay to detect Cryptosporidium spp., C. belli and C. cayetanensis simultaneously from a stool sample and described it here as coccidian mPCR. METHODS: New primers for C. belli and C. cayetanensis were designed and uniplex PCRs were standardized. The coccidian mPCR was standardized with known positive DNA control isolates. It was validated with 58 known positive and 58 known negative stool samples, which were previously identified by uniplex PCR. RESULTS: The coccidian mPCR was standardized with earlier primers designed by us for Cryptosporidium spp. and C. cayetanensis, and a newly designed primer for the internal transcribed spacer-1 (ITS-1) gene for C. belli. The coccidian mPCR was 92.1% sensitive for Cryptosporidium spp., and 100% sensitive for C. belli and C. cayetanensis each, when tested on 116 known samples. It was 100% specific for all intestinal coccidian parasites. Two representative PCR products of the newly designed ITS-1 primer for C. belli were sequenced and submitted to the GenBank, which best match with the sequences of C. belli. CONCLUSION: A highly sensitive, specific, cost-effective, indigenous, single-run coccidian mPCR has been developed, which can simultaneously detect Cryptosporidium spp., C. belli and C. cayetanensis.

Soil-Transmitted Helminth Infections among Antenatal Women in Primary Care Settings in Southern India: Prevalence, Associated Factors and Effect of Anti-Helminthic Treatment.

Community-based studies from India on prevalence of soil-transmitted helminth (STH) infections have reported estimates as high as 50% in children. However, prevalence estimates during pregnancy in India are lacking. We aimed to describe the burden, associated factors of STH and cure rate after deworming in primary care settings. Pregnant women were recruited from four urban and five rural centers in Puducherry, South India, from December 2019 to April 2022. One stool sample was collected from each participant before deworming and one repeat sample was collected from STH positive woman after three weeks of deworming. The samples were processed with saline; iodine wet mount, and microscopic concentration techniques. Cure rate (CR) was assessed using Kato-Katz thick smear. Of 650 women included, 49 (7.5%, 95% CI 5.6-9.8) had one of the STH infections; the prevalence of Ascaris lumbricoides, hookworm and Strongyloides was 5.4%, 1.8% and 0.3%, respectively. The prevalence of any STH was higher among ages 26-30 years (9.1%), working women (8.3%), multigravida (8.3%), urban setting (8.3%), those who did not wash their hands before food (9%) and anemic women (8.9%), compared to their counterparts, but not statistically significant. The CR for hookworm was 100% and Ascaris lumbricoides was 88.6%. To conclude, the prevalence of STH was low among pregnant women compared to school aged children. Continued deworming activities along with improved sanitation could further reduce the burden.

Assessment of Plasmodium falciparum histidine rich protein 2 and /3 (pfhrp 2&/ pfhrp 3) gene deletion or mutation in Plasmodium falciparum positive blood samples in a tertiary care centre in South India.

Rapid diagnostic card tests (RDTs) enable timely and appropriate diagnosis of malaria especially in remote areas. Plasmodium falciparum histidine rich protein 2 (PFHRP2) is the most targeted antigen for the detection of Plasmodium falciparum infections by rapid diagnostic card test. Genetic mutations and gene deletions are important emerging factors for false-negative RDTs, which may delay the provision of life-saving treatment for the patients. Hence, we would like to evaluate for the existence of pfhrp2/3 gene deleted P. falciparum parasites in our health care setting. This study was conducted for a period of 2 years in a tertiary care centre in South India. Blood samples that are microscopically confirmed as P. falciparum but negative by RDT were assessed for the presence of pfhrp2, pfhrp3, and their flanking genes using conventional PCR. Follow up of the clinical outcomes were also done for these patients. Of the 63 positive samples collected (50 /63) 79.4% were P.vivax and (13/63) 20.6% were P.falciparum by PCR. Among the 13 P. falciparum positive samples, 4 samples (4/13), (95% CI -10.36% to 61.11%) were found to be RDT negative but microscopically positive.Pfhrp2,pfhrp3 and their flanking genes were amplified for these 4 samples. All 4 samples were found to be negative for both pfhrp2-2 & pfhrp2-3 exon regions and also varying patterns of flanking gene deletions were also noted.This study provides molecular evidence for the existence of pfhrp2 & pfhrp3 deleted P. falciparum parasites in a tertiary care centre in South India warranting periodic evaluation of pfhrp2 based RDT use. Only pfhrp2/3 RDT based decision on diagnosis of P.falciparum malaria should always be reconsidered especially in remote areas.

Molecular characterization of Giardia intestinalis assemblages in children among the rural and urban population of Pondicherry, India.

Introduction: Giardiasis is one of the greatest public parasitic infections causing diarrheal and also known to be associated with high morbidity and mortality, among the children's particularly in developing countries with less cleanliness practices. Thus, studying genomic variety of Giardia intestinalis aids to improve our perspective related to the variability in the genome of the parasite. Materials and Methods: In this cross-sectional study, 1006 stool samples were collected from the rural (n = 500) and urban settings (n = 506) from the children (<15 years) with and without symptoms and were screened for the presence of G. intestinalis by polymerase chain reaction (PCR) targeting triosephosphate isomerase gene. Further, all PCR-positive amplicons were subjected to restriction fragment length polymorphism using RsaI restriction enzyme. Results: Of the total 1006 stool samples, 500 samples from rural screened by PCR 108 (21%) were found to be positive for assemblage A, 116 (23.2%) belong to assemblage B, and 5 (1%) were mixed assemblages (A + B). Whereas in urban, of the 506 samples screened by PCR, 92 (18.1%) were found to be positive for assemblage A, 93 (18.3%) assemblage B, and 10 (1.9%) were mixed assemblages (A + B). No significant difference was found between the G. intestinalis assemblages with clinical details of symptomatic and asymptomatic in children. Conclusions: This signifies the first study inspection in our location to shed lights and delivers some preliminary data on assemblages and subassemblages. The results suggest that anthroponotic transmission could be a foremost transmission path for giardiasis among the study population.

A Comparative Analysis of Microscopy, Coproantigen Serology, and Nested Multiplex PCR in the Laboratory Diagnosis of Entamoeba histolytica Infection.

Objectives Amoebiasis is caused by the most common intestinal protozoan parasite Entamoeba histolytica . This parasite causes amoebic colitis, which is manifested by diarrhea, followed by dysentery. The laboratory diagnosis of intestinal amoebiasis in most cases is by microscopic examination of stool samples. Other nonroutine methods include coproantigen enzyme-linked immunosorbent assay (ELISA) from stool samples, serum ELISA for antibodies, stool culture, isoenzyme analysis, and polymerase chain reaction (PCR). The present study aimed to comparatively analyze the different diagnostic modalities used for the detection of E. histolytica from the stool sample of patients with intestinal amoebiasis. Materials and Methods This study was undertaken with 631 patients, during a period of 3 years, from January 2017 to December 2019. Stool specimen obtained from each patient was subjected to direct microscopic wet mount examination, coproantigen ELISA, and nested multiplex PCR, respectively. Results Out of all the patients tested, 5.2% were positive for E. histolytica. Among the positive cases, stool microscopy was positive in 3.17%, coproantigen ELISA was positive in 29 (4.6%) cases, and PCR was positive in 30 (4.75%) cases. Statistical Analysis The prevalence of E. histolytica infection was summarized as percentages. The three diagnostic tests done were statistically analyzed, taking microscopy as the gold standard. The agreement between techniques (microscopy, coproantigen ELISA, and PCR) was analyzed with kappa statistics. Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy were summarized as percentage with 95% confidence interval. Conclusion In all suspected amoebiasis cases, a combination of stool microscopy, coproantigen testing with molecular detection of the parasite offers the best approach to diagnosis of this parasitic infection.

Detection of Entamoeba histolytica and bacterial etiological agents in patients with clinically suspected cases of liver abscesses.

Liver abscess is one of the conditions having multiple etiological agents. It can be parasitic or can be due to bacterial among other causes. Entamoeba histolytica is one of the common protozoan parasites causing amoebic liver abscess. So, accurate diagnosis is important for proper management and treatment. We have tried to detect the various bacterial etiological agents along with Entamoeba histolytica using culture of bacteria and polymerase chain reaction for E. histolytica in suspected liver abscess cases. Liver aspirates/pus collected from 63 patients were subjected to bacterial gram staining and culture along with wet mount and PCR for E. histolytica. Patients' clinical details and outcomes were also noted and co-related.It was seen that 22 (34.9%) out of 63 samples showed the presence of bacteria by gram staining whereas aerobic bacterial growth was seen in 28.6% and only 1.6% in anaerobic culture. Amoebic liver abscess showed E. histolytica  in 36 patients out of 63 study participants (57.1%) by PCR. The study showed that 44.4% of patients had a habit of alcohol consumption and 19.1% were chronic smokers. Abdominal pain (90.3%) was the most common presenting feature followed by fever (64.5%). The most common co-morbidities in the enrolled patients was diabetes mellitus (19.3%) and least with chronic liver disease (3.2%). Liver abscess, a multi-etiological condition needs a robust diagnostic method. Just a single method or a single sample type is not sufficient to diagnose, as it may miss out other causes. Treating its associated co-morbidities may help to lessen it.

Prevalence of intestinal parasitic infection and its associated factors among children in Puducherry, South India: a community-based study.

There is paucity of studies at community level on prevalence of intestinal parasitic infection among under 18 years age group. This cross-sectional community-based research aimed to determine the prevalence of intestinal worm infections and its associated risk factors among 1 to 18 years age group in Puducherry, India. Sociodemographic, behavioral and other associated factors were collected using a structured questionnaire. One stool sample was collected from each participant and examined using direct (saline/iodine wet mount) and concentration (floatation/sedimentation) microscopic techniques. Log binomial regression analysis was used to find the factors independently associated with intestinal parasitic infection. Of 187 participants who provided the stool sample, 25 (13.4%) had at least one of the parasitic infections and among them 12 (6.4%) had Soil Transmitted Helminth infection (STH) and 13 (6.9%) had intestinal protozoan parasites. Parasitic infection is marginally higher among 1 to 7 years age group (14.4%) compared to 8 to 18 years age group (12.1%). After adjusting for confounding, urban residence (APR = 3.3, 95% CI 1.4-8.0) and open-air defecation (APR = 3.3, 95% CI 1.4-7.5) were significantly associated with intestinal parasitic infections. One out of eight children had any of the parasitic infection and nearly 50% of parasitic infections were caused by STH. Those children residing in urban areas and practice of open-air defecation had higher prevalence of parasitic infection.

Tuberculosis-Learning the Impact of Nutrition (TB LION): protocol for an interventional study to decrease TB risk in household contacts.

BACKGROUND: Comorbidities such as undernutrition and parasitic infections are widespread in India and other tuberculosis (TB)-endemic countries. This study examines how these conditions as well as food supplementation and parasite treatment might alter immune responses to Mycobacterium tuberculosis (Mtb) infection and risk of progression to TB disease. METHODS: This is a 5-year prospective clinical trial at Jawaharlal Institute of Post Graduate Medical Education and Research in Puducherry, Tamil Nadu, India. We aim to enroll 760 household contacts (HHC) of adults with active TB in order to identify 120 who are followed prospectively for 2 years: Thirty QuantiFERON-TB Gold Plus (QFT-Plus) positive HHCs ≥ 18 years of age in four proposed groups: (1) undernourished (body mass index [BMI] < 18.5 kg/m2); (2) participants with a BMI ≥ 18.5 kg/m2 who have a parasitic infection (3) undernourished participants with a parasitic infection and (4) controls-participants with BMI ≥ 18.5 kg/m2 and without parasitic infection. We assess immune response at baseline and after food supplementation (for participants with BMI < 18.5 kg/m2) and parasite treatment (for participants with parasites). Detailed nutritional assessments, anthropometry, and parasite testing through polymerase chain reaction (PCR) and microscopy are performed. In addition, at serial time points, these samples will be further analyzed using flow cytometry and whole blood transcriptomics to elucidate the immune mechanisms involved in disease progression. CONCLUSIONS: This study will help determine whether undernutrition and parasite infection are associated with gene signatures that predict risk of TB and whether providing nutritional supplementation and/or treating parasitic infections improves immune response towards this infection. This study transcends individual level care and presents the opportunity to benefit the population at large by analyzing factors that affect disease progression potentially reducing the overall burden of people who progress to TB disease. Trial registration ClinicalTrials.gov; NCT03598842; Registered on July 26, 2018; https://clinicaltrials.gov/ct2/show/NCT03598842.

Molecular detection of Cystoisospora belli by single-run polymerase chain reaction in stool samples.

INTRODUCTION: Cystoisospora belli (C. belli) is the only pathogenic species of the Cystoisospora genus responsible for severe diarrhea in immunocompromised patients. Most common microscopic method of diagnosis is less sensitive due to intermittent shedding of oocysts. We developed a new single-run polymerase chain reaction (PCR)-based diagnostic assay for C. belli. METHODS: A new single-run PCR-based diagnostic assay was standardized for the detection of C. belli. Diagnostic reproducibility and repeatability of the PCR assay were evaluated. A cross-sectional analytical study was done on a total of 354 stool samples collected from 331 immunocompromised patients with diarrhea. All the stool samples were tested for the presence of oocysts of C. belli and were also tested by our new PCR assay for C. belli. Three of the representative PCR products were confirmed by sequencing. Fisher's exact test was used to compare the two proportions. RESULTS: Microscopy detected C. belli in 11/354 (3.1%) of stool samples, and the new PCR-based assay detected C. belli in 16/354 (4.5%). The new single-run PCR-based assay detected C. belli in all the stool samples which were tested positive by microscopy and additionally detected C. belli in five stool samples. The developed PCR assay detected statistically significant proportion of C. belli (p < 0.001) as compared to microscopy. The 795 base pair PCR product from one microscopy positive stool sample and two microscopy negative stool samples were confirmed by sequencing. CONCLUSION: Our newly developed single-run PCR-based detection assay for C. belli is robust and reproducible. It may be used for molecular diagnosis of cystoisosporiasis especially in transplant, pediatrics, and human immunodeficiency virus (HIV) positive patients.

Cryptosporidium and waterborne outbreaks - A mini review.

Cryptosporidium spp. is one of the prime agents of infectious diarrhea. Cryptosporidium spp. has been gaining awareness as a pathogen of public health importance in India and other developing countries. Owing to the nature of multiple transmission routes such as person-to-person, animal-to-person, waterborne and foodborne, the epidemiology of cryptosporidiosis in humans is not well known. A deeper understanding of the pathogenesis may lead to better diagnosis and better treatment of the condition. Asymptomatic human and animal transmission illustrates that the spread of infection through the environment is a more plausible explanation, waterborne transmission in particular. The disease burden is underestimated and its global impact is yet to be quantified due to the lack of country-specific estimates. Assessment of the disease itself has been crucial since the morphological indistinguishability, differences in distribution and transmission, and variations in the genotypes.

Pulmonary cryptosporidiosis in a case of adenocarcinoma of stomach: A rare case report.

Cryptosporidium species are commonly known to cause chronic intractable diarrhea in patients suffering from human immunodeficiency virus (HIV)-acquired immunodeficiency syndrome, however extra-intestinal presentations have been rarely reported. Hereby, we report a rare case of isolated pulmonary cryptosporidiosis in a 75-year-old HIV-negative patient with metastatic carcinoma of the stomach who was managed conservatively with hemostatic radiotherapy for palliative care. The patient had presented with cough with expectoration for 2 months. Sputum microscopic examination was suggestive of pulmonary cryptosporidiosis. There was no evidence of intestinal cryptosporidiosis. Therapy for pulmonary cryptosporidiosis was started with tablet nitazoxanide. The patient succumbed to the disease few days later following discharge. Although rare, patients with disseminated gastrointestinal malignancy can potentially have isolated pulmonary cryptosporidiosis.

Intestinal parasitic infections and its trends: a 5-year findings from a tertiary care centre, Puducherry, South India.

Intestinal parasitic infection (IPI) constitute a global health burden causing clinical morbidity in 450 million people. Many of these are women of reproductive age and children in developing countries. Mass deworming programmes with improvement in lifestyle are likely to reduce the intensity and prevalence of infection over the years. Hence, we aimed to assess the prevalence of intestinal parasitic infections among patients in a tertiary healthcare setting and to examine its time trends. A descriptive cross-sectional study was done using routinely collected data in a tertiary care hospital in South India. Details of examination of stool samples for the presence of intestinal helminth and protozoan ova/cysts, over the period of 5 years (2014-2019) were extracted from laboratory register and hospital information system. The presence of intestinal parasitic infection was determined by stool microscopy (direct wet mount and concentration techniques). Of the total 3267 stool samples, 303 (9.3%) had at least one parasite; 3.9% (93/3267) with helminths and 2.5% (81/3267) Entamoeba and multi-parasitism was seen in 0.14%. Stool samples from more than 18 years age had high positivity rate than others. Majority of the helminth infections were caused by Ascaris (57%) followed by hookworm (42%). Initially IPI which was 10.9% in 2014 declined to 10% in 2016 and attained a peak of 12.4% in 2017 then decreased to 6.7% in 2018. Nearly one out of ten patients had a parasitic infection. Prevalence surveys in the community followed by strengthening the deworming procedures will reduce the burden of IPIs.

'I was scared I will end up in another abortion': a mixed-methods study assessing the impact of COVID-19 pandemic and lockdown on the antenatal care of pregnant women in Puducherry, South India.

BACKGROUND: As a mitigation measure for COVID-19 pandemic, lockdown was implemented in India for a period of 2 months (24 March-31 May 2020). Disruption in antenatal care (ANC) provisions during lockdown is expected due to diversion of public health facilities on pandemic. OBJECTIVE: To assess the proportion of pregnant women who had not completed the ideal number of antenatal visits, availability of iron-folic acid (IFA) supplements and challenges in availing health services during the period of lockdown. METHODS: A concurrent mixed-methods study was conducted among pregnant women in Puducherry, India. Information on obstetric characteristics and details regarding antenatal visits were collected through telephonic interviews. In-depth interviews were conducted to understand the perceived challenges in availing health services during the lockdown period. RESULTS: Out of 150 pregnant women, 62 [41.3%; 95% confidence interval (CI) 33.6-49.3] did not complete the ideal number of visits and 61 (40.7%, 95% CI 32.7-49.0) developed health problems. Out of 44 women who received medical care for health problems, 11 (25%) used teleconsultation. Of all the women, 13 (8.7%, 95% CI 4.9-14.0) had not taken the IFA supplements as prescribed by the health provider. Economic hardship, restricted mobility, lack of information about the health system changes and psychological stress due to the fear of COVID were the challenges in accessing care. CONCLUSIONS: Two out of five pregnant women did not complete the ideal number of visits and developed health problems during the lockdown period.

Comparision of the various routine diagnostic modalities of malaria and a new method: the Parasight™ platform.

Malaria is one of most important parasitic disease, which is still much prevalent in India. The burden of malaria in India is complex and the proportions of Plasmodium vivax and Plasmodium falciparum vary across India, because of the highly variable malaria eco-epidemiological profiles, transmission factors, and the presence of multiple Plasmodium species and Anopheles vectors. The diagnostic modalities which were being used currently, are at the risk of missing potential malaria cases, if a single test is being used for a given sample. There are some extremely sensitive and specific diagnostic methods available (e.g. PCR, LAMP), but they are expensive, complex, and not readily available in all healthcare setups. Therefore, this study aimed to compare three different types of routinely used diagnostic methods and a novel testing method, the Parasight™ platform, and compare them with the detection ability of the most accurate diagnostic method, that is, PCR. A total of 111 consecutive malaria-positive (proven positive by PCR) patients were taken and tested by the immunochromatographic test or the rapid diagnostic test (RDT), thin and thick blood smears, quantitative buffy coat (QBC). In the last year of study period, 26 PCR positive samples were also taken up for the Parasight™ platform diagnostic test, along with the other routine tests. Among 111 PCR-positive cases, 78.4% samples were positive by Giemsa-stained blood film examination, 80.2% by QBC, 87.4% by RDT. In the last year of study period, among the 26 PCR-positive malaria samples, 80.8% were positive by blood film examination, 84.6% by QBC, 96.2% by RDT and 100% by the Parasight™ platform test. A combination of tests is preferable than a single method, for better detection of Plasmodium species including automated methods. The new testing method, the Parasight™ platform, is emerging to be a very sensitive test for detection of Plasmodium spp., results of which are comparable to PCR.

A comparative study assessing the effect of haematological and biochemical parameters on the pathogenesis of malaria.

Malaria is one of the most common parasitic disease affecting mankind since millennia. The most pronounced changes related to malaria involve the blood and the blood forming system, the spleen and the liver. The abnormal haematological and biochemical parameters observed in malaria cases adversely affect the prognosis of the disease. The aim of this study was to assess the severity of malaria by observing the significant abnormalities in haematological and biochemical parameters in the malaria infected cases as compared to the healthy controls. The study population comprised of 138 individuals, of which 69 were malaria cases and 69 were apparently healthy controls. All the 138 individuals were subjected to haematological and biochemical workup, following which statistical analysis was done to observe any association of altered haematological and biochemical parameters with severity of malaria, as compared to the healthy controls. Among the 138 study population, 69 patients were malaria cases whereas the other 69 were healthy controls. Haematological investigations revealed, that the haemoglobin levels, total RBC counts and haematocrit were significantly altered in the malaria cases as compared to the healthy controls. Also the leucogram profile showed significant leucopenia and neutropenia in the malaria patients as compared to the controls. Thrombocytopenia was also seen to be more pronounced in the malaria infected. The liver enzymes and serum bilirubin levels were raised in the malaria cases more than the controls. Altered haematological and biochemical parameters are indicators of disease progression to severity. Early detection and management of these parameters, will prevent the development of complications in malaria.

Genetic diversity of Indian Plasmodium vivax isolates based on the analysis of PvMSP3ß polymorphic marker.

BACKGROUND: Malaria is one of the major communicable diseases in India and worldwide. PvMSP3ß is a highly polymorphic gene due to its large insertions and deletions in the central alanine-rich region, which, in turn, makes it a valuable marker for population genetic analysis. Very few studies are available from India about the genetic diversity of Plasmodium vivax based on PvMSP3ß gene, and hence, this study was designed to understand the molecular diversity of the P. vivax malaria parasite. The accumulating epidemiological data provide insights into the circulating genetic variants of P. vivax in India, and ultimately benefits the vaccine development. MATERIALS AND METHODS: A total of 268 samples confirmed to be positive by microscopy, rapid diagnostic test, and quantitative buffy coat test were collected from four different regions of India (Puducherry, Mangaluru, Jodhpur, and Cuttack) in the present study. Polymerase chain reaction (PCR)-based diagnosis was carried out to confirm the P. vivax monoinfection, and only the mono-infected samples were subjected to PvMSP3ß gene amplification and further restriction fragment length polymorphism (RFLP) to determine suballeles. RESULTS: Based on the size of the amplified fragment, the PvMSP3ß gene was apportioned into two major types, namely Type A genotype (1.6-2 Kb) was predominantly present in 148 isolates and Type B (1-1.5 Kb) was observed in 110 isolates. The percentage of mixed infections by PCR was 3.73%. All the PCR products were subjected to RFLP to categorize into suballeles and we detected 39 suballeles (A1-A39) in Type A, and 23 suballeles (B1-B23) in Type B genotype. A high degree of diversity was observed among the isolates collected from Mangaluru region when compared to isolates collected from other regions. CONCLUSION: The present study showed a high degree of genetic diversity of PvMSP3ß gene among the isolates collected from various parts of India. High polymorphism in PvMSP3ß gene makes it a promising marker for epidemiological and vaccine development studies.