PCR-RFLP Based genetic diversity of Plasmodium vivax genotypes in district Mardan, Pakistan / Diversidade genética baseada em PCR RFLP de genótipos de Plasmodium vivax no distrito de Mardan, Paquistão

Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3α and Pvmsp-3βgenes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3α and Pvmsp3β genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp 3α genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A: 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3βgenes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3α and Pvmsp-3β genes of P. vivax isolates by using PCR/RFLP from District Mardan and [...].

O Plasmodium vivax é o parasita da malária humana mais comum nos países asiáticos, incluindo o Paquistão. O presente estudo foi desenhado para explorar a diversidade genética de genótipos de Plasmodium vivax baseados nos genes Pvmsp-3α e Pvmsp-3β, usando marcadores de ensaios alélicos nested PCR e RFLP de isolados de campo no distrito de Mardan, Paquistão. Amostras de sangue de 200 pacientes com malária por P. vivax foram coletadas após assinatura do termo de consentimento livre e esclarecido. A diversidade genética em produtos de PCR nested foi determinada por polimorfismo de fragmento de restrição (RFLP) utilizando as enzimas de restrição Alu1 e PstI para a digestão dos produtos dos genes alfa e beta, respectivamente. Para análise da diversidade genética das variantes subalélicas dos genes Pvmsp3α e Pvmsp3β, o teste Qui-quadrado foi realizado utilizando o software de programação Minitab 18. O valor P = 0,05 foi considerado estatisticamente significativo. Para os genes Pvmsp 3α, após eletroforese em gel de produtos digeridos, quatro genótipos distintos foram obtidos de um total de 50 amostras; tipo A: 35 (70%) (1,5-2,0 kb), 12 do tipo B (24%) (1,5-1,7 kb), 2 do tipo C (4%) (0,5-1,5) e um para o tipo D (2%) (0,5-0,65 kb), que podem ser caracterizados em nove padrões alélicos (A1-A4, B1-B3, C1, D), em que A3 permaneceu como o mais predominante. Para Pvmsp-3βgenes, três genótipos distintos foram obtidos a partir de 50 amostras; 40 (80%) do tipo A (1,5-2,5 kb), 9 (18%) do tipo B (1,0-1,5 kb) e 1 (2%) do tipo C (0,65 kb), que podem ser caracterizados em seis padrões alélicos (A1-A3, B1-B2 e C1). Os mais dominantes no tipo A foram o alelo A1, observados em 46%, enquanto, no tipo B, os mais dominantes foram B1 (10%). Este estudo é o primeiro relato de epidemiologia molecular e variação genética em Pvmsp-3α. Os genes Pvmsp-3β de isolados de P. vivax utilizando PCR/RFLP do Distrito Mardan mostraram um nível notável de diversidade genética nos genes estudados [...].

PCR-RFLP Based genetic diversity of Plasmodium vivax genotypes in district Mardan, Pakistan

Abstract Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3 and Pvmsp-3genes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3 and Pvmsp3 genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp-3 genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A: 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3genes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3 and Pvmsp-3 genes of P. vivax isolates by using PCR/RFLP from District Mardan and showed a remarkable level of genetic diversity in the studied genes of circulating parasites in the study area. The results of this study will contribute in future studies about the genetic structure of parasite and vaccine development against the malaria.

Resumo O Plasmodium vivax é o parasita da malária humana mais comum nos países asiáticos, incluindo o Paquistão. O presente estudo foi desenhado para explorar a diversidade genética de genótipos de Plasmodium vivax baseados nos genes Pvmsp-3 e Pvmsp-3, usando marcadores de ensaios alélicos nested PCR e RFLP de isolados de campo no distrito de Mardan, Paquistão. Amostras de sangue de 200 pacientes com malária por P. vivax foram coletadas após assinatura do termo de consentimento livre e esclarecido. A diversidade genética em produtos de PCR nested foi determinada por polimorfismo de fragmento de restrição (RFLP) utilizando as enzimas de restrição Alu1 e PstI para a digestão dos produtos dos genes alfa e beta, respectivamente. Para análise da diversidade genética das variantes subalélicas dos genes Pvmsp3 e Pvmsp3, o teste Qui-quadrado foi realizado utilizando o software de programação Minitab 18. O valor P = 0,05 foi considerado estatisticamente significativo. Para os genes Pvmsp-3, após eletroforese em gel de produtos digeridos, quatro genótipos distintos foram obtidos de um total de 50 amostras; tipo A: 35 (70%) (1,5-2,0 kb), 12 do tipo B (24%) (1,5-1,7 kb), 2 do tipo C (4%) (0,5-1,5) e um para o tipo D (2%) (0,5-0,65 kb), que podem ser caracterizados em nove padrões alélicos (A1-A4, B1-B3, C1, D), em que A3 permaneceu como o mais predominante. Para Pvmsp-3genes, três genótipos distintos foram obtidos a partir de 50 amostras; 40 (80%) do tipo A (1,5-2,5 kb), 9 (18%) do tipo B (1,0-1,5 kb) e 1 (2%) do tipo C (0,65 kb), que podem ser caracterizados em seis padrões alélicos (A1-A3, B1-B2 e C1). Os mais dominantes no tipo A foram o alelo A1, observados em 46%, enquanto, no tipo B, os mais dominantes foram B1 (10%). Este estudo é o primeiro relato de epidemiologia molecular e variação genética em Pvmsp-3. Os genes Pvmsp-3 de isolados de P. vivax utilizando PCR/RFLP do Distrito Mardan mostraram um nível notável de diversidade genética nos genes estudados de parasitas circulantes na área de estudo. Os resultados desse estudo contribuirão em estudos futuros sobre a estrutura genética do parasita e o desenvolvimento de vacinas contra a malária.

PCR-RFLP Based genetic diversity of Plasmodium vivax genotypes in district Mardan, Pakistan / Diversidade genética baseada em PCR-RFLP de genótipos de Plasmodium vivax no distrito de Mardan, Paquistão

Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3α and Pvmsp-3ßgenes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3α and Pvmsp3ß genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp3α genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A: 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3ßgenes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3α and Pvmsp-3ß genes of P. vivax isolates by using PCR/RFLP from District Mardan and showed a remarkable level of genetic diversity in the studied genes of circulating parasites in the study area. The results of this study will contribute in future studies about the genetic structure of parasite and vaccine development against the malaria.

O Plasmodium vivax é o parasita da malária humana mais comum nos países asiáticos, incluindo o Paquistão. O presente estudo foi desenhado para explorar a diversidade genética de genótipos de Plasmodium vivax baseados nos genes Pvmsp-3α e Pvmsp-3ß, usando marcadores de ensaios alélicos nested PCR e RFLP de isolados de campo no distrito de Mardan, Paquistão. Amostras de sangue de 200 pacientes com malária por P. vivax foram coletadas após assinatura do termo de consentimento livre e esclarecido. A diversidade genética em produtos de PCR nested foi determinada por polimorfismo de fragmento de restrição (RFLP) utilizando as enzimas de restrição Alu1 e PstI para a digestão dos produtos dos genes alfa e beta, respectivamente. Para análise da diversidade genética das variantes subalélicas dos genes Pvmsp3α e Pvmsp3ß, o teste Qui-quadrado foi realizado utilizando o software de programação Minitab 18. O valor P = 0,05 foi considerado estatisticamente significativo. Para os genes Pvmsp3α, após eletroforese em gel de produtos digeridos, quatro genótipos distintos foram obtidos de um total de 50 amostras; tipo A: 35 (70%) (1,5-2,0 kb), 12 do tipo B (24%) (1,5-1,7 kb), 2 do tipo C (4%) (0,5-1,5) e um para o tipo D (2%) (0,5-0,65 kb), que podem ser caracterizados em nove padrões alélicos (A1-A4, B1-B3, C1, D), em que A3 permaneceu como o mais predominante. Para Pvmsp-3ßgenes, três genótipos distintos foram obtidos a partir de 50 amostras; 40 (80%) do tipo A (1,5-2,5 kb), 9 (18%) do tipo B (1,0-1,5 kb) e 1 (2%) do tipo C (0,65 kb), que podem ser caracterizados em seis padrões alélicos (A1-A3, B1-B2 e C1). Os mais dominantes no tipo A foram o alelo A1, observados em 46%, enquanto, no tipo B, os mais dominantes foram B1 (10%). Este estudo é o primeiro relato de epidemiologia molecular e variação genética em Pvmsp-3α. Os genes Pvmsp-3ß de isolados de P. vivax utilizando PCR/RFLP do Distrito Mardan mostraram um nível notável de diversidade genética nos genes estudados de parasitas circulantes na área de estudo. Os resultados desse estudo contribuirão em estudos futuros sobre a estrutura genética do parasita e o desenvolvimento de vacinas contra a malária.

Molecular characterization of genes encoding AmpC beta-lactamases in clinical isolates of Pseudomonas and Acinetobacter species.

Emergence of AmpC beta-lactamases in isolates of Pseudomonas and Acinetobacter species, is a threatening condition as they mediate resistance to a wide variety of β-lactam drugs, including α-methoxy-β-lactams, such as cefoxitin, narrow-, expanded- and broad-spectrum cephalosporins, aztreonam and are poorly inhibited by β-lactam inhibitor combinations. The present study was conducted to determine the occurrence of blaampC genes in these pathogenic non-fermenters for their rapid and accurate detection. Monoplex PCR was done to detect blaampC genes in 40 non-duplicate clinical Pseudomonas and Acinetobacter isolates, that were found resistant to any of the third-generation cephalosporin and cefoxitin. Multiplex PCR assay was carried out to identify family-specific AmpC beta-lactamase genes within Pseudomonas and Acinetobacter spp. PCR detected blaampC in 43.24% of Pseudomonas and 33.33% of Acinetobacter isolates. Overall 42.50% of the total isolates were found to harbour blaampC genes by PCR. By multiplex PCR, total eight (20%) isolates yielded a positive amplicon with AmpC-specific primers. High prevalence of blaampC genes in cefoxitin-resistant isolates of Pseudomonas and Acinetobacter isolates emphasizes that molecular detection methods should be carried out to know the exact prevalence of beta-lactamases.

Chronic suppurative otitis media: a clinico-microbiological menace.

Background: Chronic Suppurative Otitis Media (CSOM) is an important cause of preventable hearing loss. Global emergence of resistant strains is of great concern. The aim of the present study was to determine the etiology and antibiotic sensitivity pattern of bacterial isolates from CSOM cases with special emphasis on ESBL (Extended Spectrum Beta- Lactamases) and AmpC beta lactamases. Methods: Patients with sign and symptoms suggestive of CSOM, ESBL (Extended Spectrum Beta-Lactamases), AmpC beta lactamases and MBLs (Metallo beta lactamases) were included. Two ear swabs were taken from all the patients and cultured on blood agar and MacConkey agar. Bacterial identification of isolates was done using standard biochemicals. Antimicrobial susceptibility was performed by Kirby-Bauer's disc diffusion method as per the Clinical Laboratory Standards Institute (CLSI) guidelines using antibiotic discs (HI MEDIA). Results: Out of 130 patients, 110(84.62%) had bacterial growth. The common pathogenic species were Pseudomonas aeruginosa 36(37.89%), Staphylococcus aureus 31(32.63%), Citrobacter koseri 9(9.47%) and Proteus vulgaris 6(6.32%). P. aeruginosa showed maximum sensitivity to colistin (94.4%), polymixin-B (91.3%) and imipenem (91.3%). Gram positive cocci showed maximum sensitivity to vancomycin (99%). MRSA (Methicillin Resistant Staphylococcus aureus) and HLAR (High Level Aminoglycoside Resistance) were detected in 9(29%) S. aureus and 1(50%) Enterococcus faecalis respectively. ESBL and AmpC were detected in 11(18.3%) and 12(20%) Gram negative bacteria, respectively and MBL producer was not detected. Conclusion: P. aeruginosa was found to be the most common isolate in CSOM cases and colistin, polymixin-B and imipenem was found to be most effective antibiotics.

Increasing antimicrobial resistance among uropathogens: is fosfomycin the answer?

Urinary tract infection [UTI] is one of the most common infectious diseases in clinical practice. The choice of antibiotics for the treatment of UTI is limited by the rising rates of antibiotic resistance. There is an urgent need to discover new effective treatment solutions. Fosfomycin may be an interesting alternative to the currently used treatments of UTIs. The study was conducted over 6 months period [January to June 2013] in Department of Microbiology, JNMCH, AMU, Aligarh. A total of 1840 urine samples were submitted. Culture and sensitivity was done as per standard microbiological procedures. Methicillin-resistant Staphylococcus aureus [MRSA], high-level aminoglycoside resistance [HLAR], extended spectrum beta-lactamases [ESBL], AmpC and metallo-beta-lactamases [MBL] production was detected. Culture was positive in 504 [27.4%] cases. Gram-negative etiology was identified in 390 [73%] cases. ESBL production was detected in 154 [37.1%] while 82 [21.6%] were Amp C. No, MBL was detected. Among Gram-positive bacteria, 68 [51.5%] were MRSA, while 4 [13.3%] were vancomycin resistant enterococci [VRE]. HLAR was seen in 53.3% of enterococci. Fosfomycin was effective in 100% of MRSA, VRE, ESBL, HLAR, and overall, susceptibility to fosfomycin in AmpC producers was extremely high [99%]. Norfloxacin and cotrimoxazole were not proved effective as only three isolates were sensitive to norfloxacin, while all Gram-negative isolates were resistant to cotrimoxazole. Pseudomonas species showed 65% and 75% susceptibility to colistin and polymixin B, respectively. Fosfomycin has emerged as a promising option, especially in cases involving multi-drug-resistant pathogens in which previous antibiotics have failed to cure the infection

IgG Avidity Antibodies against Toxoplasma gondii in High Risk Females of Reproductive Age Group in India

Toxoplasma gondii is an obligate intracellular protozoan that is distributed worldwide. Recently, several tests for avidity of Toxoplasma IgG antibodies have been introduced to help discriminate between recently acquired and distant infections. The study was conducted in Jawaharlal Nehru Medical College and Hospital, India from February 2011 to September 2012. Serum specimens were subjected to Toxoplasma IgM ELISA and IgG avidity ELISA test. Out of 48 patients with abortions, 17 (35.4%) were positive for IgM ELISA, and 8 (16.6%) had low IgG avidity antibodies. Out of 48 patients with other obstetric problems, 23 (47.9%) were positive for IgM ELISA, and 17 (35.4%) had low IgG avidity antibodies. Combining both groups on avidity test, only 25 of 40 (62.5%) IgM-positive women had low-avidity IgG antibodies suggesting a recent T. gondii infection in these women. More importantly, 15 (37.5%) of the IgM-positive women had high-avidity antibodies suggesting that the infection was acquired before gestation The relation of IgM seropositivity with the following risk factors was not found to be statistically significant; contact with cats (0.13), non-vegetarian food habits (0.05), and low socio-economic status (0.49). While, for IgG avidity ELISA, only contact with cats (0.01) was significantly associated with seropositivity. All other risk factors have P-values of >0.05 (not significant). IgG avidity test when used in combination with IgM test was a valuable assay for diagnosis of ongoing or recently acquired T. gondii infection in India.

Antibacterial activity of silver nanoparticles dispersion against MSSA and MRSA isolated from wounds in a tertiary care hospital of north India.

Staphylococcus aureus, the main cause of nosocomial infection worldwide result in significant increases in mortality, morbidity, and cost related to prolong treatments. Silver compound has been in use since time immemorial for the treatment of burns, wounds and several other bacterial infections. In the present work, we explore the antibacterial activity of silver nanoparticles (Ag-NPs) dispersion (5-10 nm) against reference strain and clinical isolates of Methicillin-sensitive S. aureus (MSSA), and Methicillin-Resistant S. aureus (MRSA).The typical minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against standard reference strain as well as, MSSA and MRSA were observed in the range of 12-48 μg/ml and 12-96 μg/ml, respectively. The MBC/MIC ratios against all strains were found in the range of ≤1 to ≤4, which shows that Ag-NPs inhibit bacterial growth in a bactericidal rather than a bacteriostatic manner. Our finding suggests that Ag-NPs are effective broad-spectrum antibacterial agents regardless of their drug-resistance mechanisms.

Comparative study on occurrence of class A and class C β -lactamase genes and their co-occurrence in Indian Enterobacteriaceae during years 2009 and 2010

OBJECTIVE@#To determine the occurrence of class A and class C β-lactamase genes and their co-occurrence in Indian Enterobacteriaceae.@*METHODS@#52 third generation cephalosporin resistant isolates were phenotypically detected by combination disk method and screened by PCR to identify class A and class C type β-lactamase genes.@*RESULTS@#Of the 52 isolates, 94.2% (49) were found harboring any of the bla(ESBL(s)). bla(CTX-M), bla(SHV) and bla(TEM) were present in 82.6% (43/52), 59.6% (31/52), and 42.3% (22/52) isolates, respectively. Of the 49 ESBL positive isolates 57.1% (28/49) showed co-occurrence of bla(ampC) with bla(ESBL(s)). On the contrary, the collection from 2009 showed their co-occurrence in 81.4% isolates.@*CONCLUSIONS@#The comparative study shows a downward trend for co-existence of bla(ESBL(s)) with bla(ampC) from 2009 to 2010. Further large scale studies are needed to address the co-occurrence of class A and class C β-lactamases in India and the resistance trend occurring over a period of time.

Changing trends in prevalence of different Plasmodium species with dominance of Plasmodium falciparum malaria infection in Aligarh (India)

OBJECTIVE@#To determine the prevalence of malaria in Aligarh and analyze species dominance in different years over a decade.@*METHODS@#Diagnosis of malaria was done using microscopy as gold standard, rapid antigen detection assays and quantitative buffy coat (QBC) assays. Giemsa stained blood smear examination was done, thick and thin films were examined for presence of different Plasmodium spp. Rapid antigen detection assays employing detection of HRP-2 and parasite lactate dehydrogenase antigen (pLDH) by immunochromatography was done in patients whose blood smear found to be negative by conventional Giemsa slide examination. QBC was done in cases where there is strong clinical suspicion of malaria with blood smear negative, in patients with chronic malaria, splenomegaly, or in those patients who had inadequate treatment and for post-treatment follow up.@*RESULTS@#Plasmodium vivax and Plasmodium falciparum were only species detected in our hospital. Overall prevalence of malaria in Aligarh was found to be 8.8%. The maximum prevalence of 20.1% was observed in year 2008 and lowest 2.3% in 2002.@*CONCLUSIONS@#High prevalence of malaria is observed in this part of country with dominance of both species particularly Plasmodium falciparum should be monitored and factors accounting for occurrence should be studied to employ effective control measures.

Evaluationof ParaSight F Test in Dignosis of Plasmodium Falciparum Infection.

P. falciparum malaria is a severe form of disease which requires urgent diagnosis and treatment to save the life of patient. Blood smear examination is the commonest method used for diagnosis. The present study was done to evaluate ParaSight F test in patients of P. falciparum infection.The study was performed on 100 patients who where clinically diagnosed as cases of P. falciparum infection. ParaSight F test and Leishman stained blood smear examination was done in all 100 patients (50 patients of cerebral malaria + 50 patients of acute malaria).ParaSight F test was positive in 45 patients and blood smear positive in 28 patients of cerebral malaria. 35 patients of acute malaria were positive by ParaSight F test while blood smear was positive in 15 patients. Sensitivity, specificity, positive and negative predictive values of ParaSight F test are 95.7%, 100%, 100%, 100%, 60% in cerebral malaria and 100% each in acute malaria. ParaSight F test can be used as diagnostic tool in cases of P. falciparum infections, where blood smear is negative.

Comparison of different diagnostic techniques in Plasmodium falciparum cerebral malaria.

BACKGROUND & OBJECTIVES: Plasmodium falciparum cerebral malaria remains a major health problem in India. The efficacy of treatment of cerebral malaria lies in its early diagnosis through rapid diagnostic methods. ParaSights-F test detects HRP-2 antigen secreted by parasitised red blood cells and quantitative buffy coat assay (QBC) is examination of buffy coat for the presence of malarial parasite stained with acridine orange. This study was performed to evaluate the effectiveness of ParaSight-F test and QBC assay as diagnostic methods in the patients of cerebral malaria. METHODS: Fifty clinically diagnosed patients of cerebral malaria were included in the study. ParaSight-F test, QBC and conventional blood smear examination was done. Patients who were in coma and there were no obvious features of bacterial or viral etiology were investigated for cerebral malaria by these diagnostic methods. RESULTS: ParaSight-F test, QBC and peripheral blood smears were examined. Patients were followed-up for signs of clinical recovery. ParaSight-F test was positive in 47 patients, QBC in 46 while blood smear examination was positive in 28 cases. INTERPRETATION & CONCLUSION: Sensitivity and specificity of ParaSight-F test were found to be 96.6 and 94% while QBC showed 97.8 and 100% respectively. ParaSight-F test and QBC were found to be novel methods for diagnosis of cerebral malaria especially in the cases where diagnosis can not be made by conventional blood smear examination due to low parasitaemia. These rapid diagnostic methods help in early therapeutic intervention.

Gall-stone composition and biochemical alterations in the serum of patients with cholelithiasis

The levels of cholesterol, calcium and bilirubin were determined in the seruih and gall stones in 47 cases with cholelithiasis. The mixed stones were observed in 87.5 percent, high cholesterol stone in 8.5 percent and high bilirubin stones in 4.2 percent. Serum and stone cholesterol levels were inversely proportional and the bilirubin levels were directly proportional to each other