Trends in clinical features and severity of Plasmodium vivax malaria among children at tertiary care center in North India.

BACKGROUND: Malaria is a significant cause of morbidity and mortality in adults and children. Plasmodium falciparum is the primary cause of severe malaria, but recently Plasmodium vivax is also recognized to cause severe malaria-associated morbidity and mortality. The study focuses on determining the mortality related to severity parameters in individuals under 12 years and their critical presentation in P.vivax malaria-infected children. METHODS: A prospective cross-sectional hospital-based study was conducted at Safdarjung Hospital, New Delhi, and ICMR-NIMR, New Delhi. All clinically suspected cases were admitted for screening. Exclusion criteria (rapid malaria antigen test, microscopy and medication history) were applied to all the admitted patients (n = 221) to obtain P.vivax patients only. Patients aged ≤ 12 years were included in the study. DNA was extracted from dried blood spots and amplified by nested PCR, followed by visualization on gel electrophoresis. RESULT: A total of 221 clinically suspected cases of malaria were screened for P.vivax. After implementing various exclusion criteria, 45/221 cases were enrolled for the study, among which 44.4% (20/45) of children had the symptoms of severe malaria in terms of cerebral malaria, thrombocytopenia, anemia, pancytopenia, acute respiratory distress syndrome and hemophagocytic lymphohistiocytosis. CONCLUSION: Plasmodium vivax mono-infection can cause severe manifestation and must be treated as P.falciparum without any delay because it may lead to increased morbidity and mortality. A changing trend in clinical symptoms has shown in P.vivax which was an earlier phenomenon of P.falciparum.

Studying the disease severity in clinical isolates of Plasmodiumvivax.

Among the human malaria Plasmodium species, Plasmodium vivax is the most widespread species globally. In recent times, this historically benign species is now being recognized as also responsible for severe malaria infections in humans. Hence, a deeper insight of P.vivax immunopathogenesis in clinical patients is essential for malaria control and elimination strategies. Certain genes like vir genes, merozoite surface protein 3α genes (msp3α) and biomarkers like super oxide dismutase (SOD-1), tumor necrosis factor (TNF- α), interleukin (IL-10) are speculated to have some role in disease severity and thus can be useful as diagnostic markers. In the reported study, the clinical samples of P.vivax were genotyped for msp3α gene and cytokine analysis, expression profiling of vir genes were also carried out in these samples. A total of 84 P.vivax samples were collected (39 severe and 45 non-severe samples) and no correlation of parasitemia with severity of disease was seen in these samples (p-value = 0.38). On analysis four genotypes of msp3α were found, with type B (1.5 kb) as the predominant genotype. Cytokine analysis revealed SOD-1 and TNF-α levels to be significantly more in the severe group than in non-severe group, whereas for IL-10 no significant difference was observed between two clinical groups. The vir gene profiling revealed increased level of expression for vir-12, vir-14 related, and vir-17 like in severe group and vir-10 related gene expression was more in non-severe samples. There are multiple factors that bring phenotypic and genotypic changes in P.vivax malaria and thus, it is important to assess the potential diagnostic markers for detection of disease severity. In future, studies with more number of clinical samples should be undertaken for better insight of P.vivax disease severity.

Molecular characterization of the severe falciparum malaria with typhoid co-infection: A case report.

Malaria and typhoid co-infections can be a serious public health issue in tropical countries leading to incorrect diagnosis due to overlapping clinical presentations of malaria and typhoid and hence, causing a delay in implementing the appropriate treatment regimen for these concurrent infections. This study reports a case of six-year-old female child co-infected with severe malaria (Plasmodium falciparum) and typhoid (Salmonella typhi) diagnosed by rapid malaria antigen test (RMAT) and blood culture respectively. Further, analysis of the chloroquine resistance gene Pfcrt for the falciparum demonstrated the presence of K76T mutant allele in pfcrt gene with high IC50 (150nM) for chloroquine (CQ) drug. The present case highlights the significance of timely identification and treatment of co-infections and also provides information about the circulating P. falciparum clinical strains.

Assessing the in vitro sensitivity with associated drug resistance polymorphisms in Plasmodium vivax clinical isolates from Delhi, India.

The drug resistance of Plasmodium vivax in clinical cases remains largely unknown till date because of the difficulty in diagnosing the resistant P. vivax strains. The present study was undertaken to determine the prevalence of mutant alleles in drug resistance genes viz P. vivax multi-drug resistance (pvmdr-1), chloroquine resistance transporter (pvcrt-o), dihydrofolate reductase (pvdhfr) and dihydropteroate synthase (pvdhps) along with in vitro chloroquine (CQ) sensitivity in P. vivax clinical isolates. During August-October 2017 a total of 86 samples of the febrile patients were screened and 31 samples were found to be positive for P. vivax in Safdarjung hospital, New Delhi. Sequence genotyping of the drug resistance genes was carried out in these P. vivax samples and in vitro CQ susceptibility for 23 isolates was determined by the schizont maturation assay (SMA). The CQ inhibitory concentrations (IC50) for the clinical isolates was found to be in the range of 25.6-176.7 nM. All the 31 clinical isolates analyzed for pvmdr-1 gene, showed mutant alleles and in only two isolates novel mutations at 861 and 898 codons were observed. Sequence analysis of pvcrt-o, pvdhfr and pvdhps genes revealed wild type genotypes in all the 31 studied isolates. The presence of mutations in pvmdr-1 gene and the increase in the CQ IC50 value indicates the possibility of shift in drug tolerance where CQ with primaquine (PQ) is still the first line of treatment for P. vivax malaria in the country. The regular molecular surveillance in P. vivax would provide useful information for the policy makers of the malaria control programme.

Severe vivax malaria trends in the last two years: a study from a tertiary care centre, Delhi, India.

BACKGROUND: Plasmodium vivax, once considered benign species, is recently being recognised to be causing severe malaria like Plasmodium falciparum. In the present study, the authors report the trends in malaria severity in P. vivax among patients from a Delhi government hospital. The aim of the study was to understand the disease severity and the burden of severe vivax malaria. METHODS: A hospital based study was carried out from June 2017 to December 2018 at a tertiary care centre from Delhi, India. Patients were tested for malaria using peripheral blood smear (PBS) and/or rapid malaria antigen test (RMAT). The severe and non-severe vivax malaria categorization was done as per the WHO guidelines. Sociodemographic, clinic and paraclinical data were collected from patients and their medical records. RESULTS: Of the 205 patients, 177 (86.3%) had P. vivax infection, 22 (10.7%) had P. falciparum infection and six (2.9%) had mixed infection with both the species. Out of 177 P. vivax cases included in this study one or more manifestations of severe malaria was found in 58 cases (32.7%). Severe anaemia (56.9%), jaundice (15%) and significant bleeding (15%) were the most common complications reported in most of patients, along with thrombocytopenia. CONCLUSIONS: In this study, it is evident that vivax malaria is emerging as the new severe disease in malaria patients, a significant shift in the paradigm of P. vivax pathogenesis. The spectrum of complications and alterations in the laboratory parameters in P. vivax clinical cases also indicate the recent shift in the disease severity.

HIV-TB Cross-referral and Collaborative Strategy: 8 Years of Our Experience from An Urban Health Centre in North India.

BACKGROUND: Human immunodeficiency virus (HIV) and tuberculosis (TB) epidemics continue to fuel each other and with dual infections with these two deadly diseases on the rise, it becomes imperative to devise effective HIV-TB collaborative strategies. The present study was designed to evaluate the existing HIV-TB cross-referral mechanism at an urban health centre; to determine HIV sero-prevalence among pulmonary TB patients referred from chest clinic to the integrated counselling and testing centre (ICTC); and to evaluate the TB suspects referred from ICTC to the chest clinic for a possible TB aetiology. METHODS: The present study was a retrospective analysis of HIV-TB cross-referrals whereby a line list of all the patients referred under this strategy from January 2006 to December 2013 was retrieved and analysed. RESULTS: A total of 3726 TB cases were referred to the ICTC and 641 TB suspects were identified by ICTC counsellors and referred to the chest clinic during this period. HIV sero-prevalence among TB patients was 2.8% (106 of 3726) and TB prevalence among HIV sero-positive and sero-negative TB suspects was 9.3% (10/108) and 4.3% (9/211), respectively (p=0.07). HIV prevalence was found to be significantly higher among male (n=2024) than among female (n=1702) TB patients (4.4% versus 0.9%; p<0.0001). Only 319 of 641 (49.8%) ICTC patients referred to the chest clinic reached there. CONCLUSION: Our study highlights the strong need to scale up the integration and partnership between HIV and TB programmes for better and integrated diagnosis and care of HIV-TB co-infected patients.

Seroprevalence and characterization of Epstein-Barr virus exposure among paediatric population.

The study explored Epstein Barr Virus (EBV) exposure in 244 children using EBV-specific serology. Seroprevalence of EBV was 75-80%. Past infection and primary infection were observed in 52.04% & 8.6% respectively, whereas 23.36% showed no serological evidence of exposure to EBV. Age-stratification suggested maternal antibodies may have protected infants till 6 months of age, while the 1-3 year age group showed maximum primary infection and the 6 months to 1 year group showed the maximum susceptible group to EBV primary infection. There is a paucity of literature about EBV in India and further research is required for a better understanding of EBV pathogenesis and its clinical implications in Indian children.

A Case-Control Study of Serum and Tissue Catalase among Morphological Variants of Vitiligo.

Background: Role of oxidative stress in the pathogenesis of vitiligo. Aim: Estimation of serum and tissue catalase levels in morphological variants of vitiligo. Settings and Design: A prospective case-control study was conducted in the outpatient department of Dermatology in Safdarjung hospital. Materials and Methods: We estimated levels of serum and tissue catalase in 30 vitiligo patients and 30 matched healthy controls. Statistical Analysis: The data analysis was done in Statistical Package for Social Sciences (SPSS) version 21.0. Normality of data was tested by Kolmogorov-Smirnov test. Results: Serum and tissue catalase was lower in vitiligo patients than controls. Serum catalase was lowest in vulgaris type, whereas in the acrofacial type had lowest tissue catalase levels. Conclusion: Vitiligo patients have a generalized oxidative stress functioning at a higher pace as seen with decreased serum and tissue CAT which can well be taken as a marker of active disease and they can be helped with topical pseudoCAT preparations.

Scrub Typhus Co-Infections in a Young Boy With Varicella and Malaria: A Rare Case Report.

Scrub typhus is one of the leading causes of acute febrile illness of unknown origin in India. Though several co-infections of other vector-borne diseases have been described in the literature, few such cases have been described in children. As such, it is challenging to diagnose scrub typhus alone and becomes that more complicated when a varicella infection precedes it. This is the first reported case where scrub with varicella infection also had concomitant malaria. In such cases, prompt diagnosis and initiation of the correct drug are imperative. Here we describe a six-year-old child with a past history of varicella infection and co-infected with scrub typhus and malaria.

Pancytopenia with hemophagocytic lymphohistiocytosis in Plasmodium falciparum: A unusual presentation.

Hematological manifestations such as anemia and thrombocytopenia are known complications in malaria. Here, we report two cases presented as pancytopenia with hepatosplenomegaly and initial diagnosis kept as hematological malignancy like leukemia but later on its diagnosed as malaria-associated hemophagocytic lymphohistiocytosis which is a rare entity. The aim of this report is to draw the attention of physicians, especially in tropical countries such as India and Sub-Saharan nations to keep in mind this uncommon presentation of malaria, though the exact pathophysiological mechanism still remains obscure.

Neonatal Chikungunya: A case series.

Chikungunya virus is an RNA virus that belongs to the family Togaviridae, genus Alphavirus, transmitted by Aedes mosquitoes. The disease usually manifests as fever, arthralgia and petechial or maculopapular rash. The illness is usually self-limiting. We report a series of neonates infected with Chikungunya virus, confirmed by ELISA test, showing that viral Chikungunya can be transmitted from mother to babies and its clinical presentation is that of septicemia or meningitis.

Development of Two-Tube Loop-Mediated Isothermal Amplification Assay for Differential Diagnosis of Plasmodium falciparum and Plasmodium vivax and Its Comparison with Loopamp™ Malaria.

To strengthen malaria surveillance, field-appropriate diagnostics requiring limited technical resources are of critical significance. Loop-mediated isothermal amplification (LAMP) based malaria diagnostic assays are potential point-of-care tests with high sensitivity and specificity and have been used in low-resource settings. Plasmodium vivax-specific consensus repeat sequence (CRS)-based and Plasmodium falciparum-specific 18S rRNA primers were designed, and a two-tube LAMP assay was developed. The diagnostic performance of a closed-tube LAMP assay and Loopamp™ Malaria Detection (Pan/Pf, Pv) kit was investigated using nested PCR confirmed mono- and co-infections of P. vivax and P. falciparum positive (n = 149) and negative (n = 67) samples. The closed-tube Pv LAMP assay showed positive amplification in 40 min (limit of detection, LOD 0.7 parasites/µL) and Pf LAMP assay in 30 min (LOD 2 parasites/µL). Pv LAMP and Pf LAMP demonstrated a sensitivity and specificity of 100% (95% CI, 95.96-100% and 89.85-100%, respectively). The LoopampTM Pan/Pf Malaria Detection kit demonstrated a sensitivity and specificity of 100%, whereas LoopampTM Pv showed a sensitivity of 98.36% (95% CI, 91.28-99.71%) and specificity of 100% (95% CI, 87.54-100%). The developed two-tube LAMP assay is highly sensitive (LOD ≤ 2 parasite/µL), demonstrating comparable results with the commercial Loopamp™ Malaria Detection (Pf/pan) kit, and was superior in detecting the P. vivax co-infection that remained undetected by the Loopamp™ Pv kit. The developed indigenous two-tube Pf/Pv malaria detection can reliably be used for mass screening in resource-limited areas endemic for both P. falciparum and P. vivax malaria.

Allelic variation of msp-3α gene in Plasmodium vivax isolates and its correlation with the severity of disease in vivax malaria.

Malaria is a global socio-economic burden of which Plasmodium vivax contributes for about 70-80 million cases on an annual basis worldwide and 60-65% cases in India. Diversity observed in highly polymorphic Merozoite Surface Protein-3α (msp-3α) encoded by MSP-3 gene family, has been used efficiently for genotyping of P. vivax infection. This study aims to correlate the severity of clinical symptoms with parasite load, genotype of P. vivax and multiplicity of infection. Based on clinical symptoms classification, 31 (67.9%) out of 46 cases were found to be severe while 15 (32.6%) were non-severe and correlation of the severity of vivax infection with parasite load was not observed. Analysis of msp3-α allele genotype showed that out of 31 severe cases, 19 (61.2%) were single-clone infection cases whereas 12 (38.7%) were multi-clone infections. Similarly, out of 15 non-severe cases, 9 (60%) were single clone and 6 (40%) were multi-clone infections indicating the absence of a correlation between the multiplicity of infection and disease severity. Allele frequency observed was 65.9%, 23.4%, 23.4%, and 28.2% for allele A, B, C and D, respectively. An important finding was the greater distribution of allele D than alleles B and C, which has been reported as a rare allele otherwise. Further, of 13 cases with allele D, 76.9% (10/13) cases were severe. This study showed the absence of a correlation between the severity of clinical symptoms with parasite load and multiplicity of infection but at the same time drives a possibility of severe vivax malarial symptoms to have an association with the persistence of allele D in the population. This upon exploration can lead to the development of a target in detection of severe cases of malaria.

Clinico-laboratory findings of patients during dengue outbreak from a tertiary care hospital in Delhi.

There are outbreaks of dengue every year in India. They vary in the predominant serotype involved, clinical features and predominant laboratory findings. This study of the 2006 outbreak in Delhi highlights clinical features and laboratory parameters of dengue cases and compares the clinical features among the adult and paediatric age groups. The 2006 outbreak had higher bleeding manifestations and a greater involvement of the adult male population than in previous outbreaks. No correlation was observed between platelet count and different bleeding manifestations until they dropped <20,000 per microL. Few patients developed a florid picture before seroconversion, highlighting a greater stress on the haematological rather than immunological profile of such patients. Atypical symptoms, such as diarrhoea, menorrhagia and seizures were often the presenting complaints rather than the typical clinical features of dengue.

Early treatment failure in concurrent dengue and mixed malaria species infection with suspected resistance to artemisinin combination therapy from a tertiary care center in Delhi: a case report.

BACKGROUND: Concurrent dengue and mixed malaria infections in a single patient present with overlapping clinical manifestations which pose a diagnostic challenge and management dilemma in areas of common endemicities. METHODS: We report a case of a young male who tested positive for both Plasmodium vivax and Plasmodium falciparum along with dengue infection. He showed signs of early treatment failure to artemisinin combination therapy (artesunate with sulfadoxine+pyrimethamine). Molecular analysis for the drug resistance genes viz: chloroquine resistance (pfcrt), multidrug resistance (pfmdr-1), sulfadoxine (pfdhps), pyrimethamine (pfdhfr), and artemisinin resistance (keltch 13) was performed. RESULTS: A rise in parasitemia from <2% to 5% was observed after 3 days of treatment. Mutations in pfcrt, pfmdr-1, pfdhfr, and pfdhps genes were detected as a possible cause of treatment failure. CONCLUSION: Increased severity, overlapping symptoms, and suspected resistance to treatment warrants a multidimensional diagnostic approach and diligent therapeutic monitoring.

Onychomycosis due to rhizomucor in psoriatic patient with HIV infection.

Onychomycosis is frequently seen in dermatological clinical practice worldwide. The causative agents are usually two pathogenic groups of fungi namely, dermatophytes and yeasts of the genus Candida. In some cases, non-dermatophytic molds belonging to different genera and species may be the etiological agents. We report an unusual case of onychomycosis in an HIV-positive psoriatic patient caused by Rhizomucor pusillus, which has not been mentioned in the literature before. Our finding underline the fact that fungal species appearing as contaminants should be evaluated by proper clinical-mycological correlation to ensure an accurate diagnosis.

Routine Screening for Rubella and CMV Antibodies During Pregnancy: Is it Justifiable?

BACKGROUND: Rubella and cytomegalovirus (CMV) screening during pregnancy is routinely carried out in India. However, its value has been questioned due to the absence of clearly effective intervention. OBJECTIVES: This retrospective study evaluates the usefulness of rubella and CMV antibody screening during pregnancy. MATERIALS AND METHODS: Serum samples received from pregnant women and children were tested for rubella- and CMV-specific IgM antibodies by capture ELISA. The data were analyzed to determine the incidence of rubella and CMV infection during pregnancy and in congenital infections. RESULTS: In asymptomatic pregnant females (n = 505), rubella positivity was 3.16 % and in women with bad obstetric history (BOH) (n = 220), it was 7.72 %, while CMV positivity was 5.9 % in both asymptomatic pregnant women and in women with BOH. In children (n = 200), the overall positivity for rubella- and CMV-specific IgM antibodies was 15 and 25 %, respectively. A declining trend was observed in the incidence of both rubella and CMV infections in pregnant women and in women with BOH. In children, the incidence of congenital rubella syndrome has declined, but the incidence of CMV infection has remained almost the same in 5 years. CONCLUSION: The incidence of rubella has reduced over the past 5 years and can further be prevented by providing direct protection to women and school girls with rubella vaccines. Primary CMV infection in pregnancy is the main problem, and due to the unavailability of efficient and safe treatment, routine antenatal screening for rubella and CMV should be reserved for women with obstetric complications only.

Immunodiagnosis of dengue virus infection using saliva.

Keeping in view the complications and the case fatality associated with dengue virus, several serologic tests have been developed. However, the major drawback of these serologic tests is the need for a venous blood sample obtained by invasive venipuncture. As a noninvasive alternative, saliva provides a body fluid that contains antibodies of diagnostic importance. Hence, the detection of DEN-specific IgM and IgG antibodies in serum and saliva from 80 patients was compared. Salivary IgM antibodies were detected in 100% of the serum IgM-positive samples and in 30% of the serum samples that were negative for IgM antibodies. Salivary IgG antibodies were detected in 93.3% of the serum samples that were positive for anti-dengue IgG antibodies and in none of the serum IgG-negative cases. None of the specimens from the healthy controls showed the presence of IgM or IgG antibodies. The detection of both IgG and IgM antibodies in saliva correlated well with the serum IgG and IgM detection by the ELISA test (r = 0.6322 and r = 0.4227). Detection of salivary IgM antibodies by ELISA showed 100% sensitivity, 70% specificity, 90.9% positive predictive value, and 100% negative predictive value. The detection of IgG in saliva proved to be a promising tool as the sensitivity, specificity, positive predictive value, and negative predictive value were found out to be 93.3%, 100%, 100%, and 83.3%, respectively. Thus, from this study we conclude that the detection of DEN-specific salivary IgG and IgM antibodies are useful markers for dengue infection.