Medical Immunology

Evaluation of reliability of full hemogram and urinalysis in relation tourine culture as diagnostic tools for urinary tract bacterial infections in Trans-Nzoia county referral hospital, Kenya

2024-12-03T13:50:15Z

Evaluation of reliability of full hemogram and urinalysis in relation tourine culture as diagnostic tools for urinary tract bacterial infections in Trans-Nzoia county referral hospital, Kenya KIPKOECH, Julius Serem In routine clinical practice, urinary tract bacterial infections (UTBIs) are diagnosed using urinalysis assay and urine culture is used as a confirmatory test to determine the infecting bacteria. Urinalysis assay relies on the levels of leukocytes, proteins, nitrites and red blood cells in the urine sample. However, in public and private hospitals without blood and urine culture tests, the levels of neutrophils and monocytes from full hemogram assay are commonly interpreted as indications of bacterial infections upon which treatment is prescribed. Prescription of broad-spectrum antibiotics based on such non-specific diagnosis may result in multi drug resistance, human suffering and unnecessary expenses. The general objective of the study was to evaluate the reliability of full hemogram assay and urinalysis assay in relation to urine culture as diagnostic tools for urinary tract bacterial infections at Trans-Nzoia County Referral Hospital in Trans-Nzoia County, in Kenya. The specific objective of this cross-sectional study was to; determine the correlation between counts of neutrophils and monocytes from full hemogram assay, determine the correlation of counts of leukocytes, proteins, nitrites and red blood cells from urinalysis assay and determine the extent of reliability of counts of full hemogram assay and urinalysis assay in relation to urine culture test. The study obtained Informed Consent from the participants prior to participation and through Simple random sampling technique, 173 participants were recruited from both in-patient and out-patient departments at the hospital. About 2-4ml of urine sample was obtained from the patients for urinalysis and urine culture tests. Urine culture in this case was used as a gold standard diagnosis for urinary tract bacterial infections. About 1ml of blood was drawn by venipuncture from the same study participants for full hemogram assay to determine the counts of monocyte and neutrophils. Out of 173 study patients, 122 patients were confirmed positive for urinary tract bacterial infections by urine culture test. Out of these positive cases, 55 patients were neutrophilic, 24 patients presented with monocytosis and 39 patients had leukocytosis. On contrast, 51 samples tested negative by urine culture and were used as control group in the study. The study used a desired confidence level of 95% and an acceptable margin error of 5%. There was no correlation between the counts of neutrophils and monocytes from full hemogram assay with value of r=0.0794 and p-value of 0.299.However, there were correlations between the variables of urinalysis assay and their means had a significant relationship with a p-value of <0.0001on one way Analysis of Variance test. Urinalysis assay was not reliable tool with an overall score of 0.17 on Cronbach's Alpha scale while full hemogram was not reliable tool with the best score of 45% in relation to urine culture test. On ranking of reliability of the variables from both assays, Neutrophils were most reliable with 45.08%, Leukocytes 31.97%, Proteins 20.49%, Monocytes 19.67% and RBCs at 9.84% in relation to confirmatory urine culture technique. In conclusion, full hemogram assay and urinalysis assay are not reliable tools for diagnosis of urinary tract bacterial infectionsin relation to the confirmatory test of urine culture technique. From the study outcome, it is recommended that hospitals should use urine culture techniques in diagnosis of urinary tract bacterial infections. Master's Thesis

Performance of one-step spanbio™ rapid test in detection of giardia lamblia in Busia county referral hospital, western Kenya

2022-12-20T16:13:31Z

Performance of one-step spanbio™ rapid test in detection of giardia lamblia in Busia county referral hospital, western Kenya INDIEKA, Roberts. Briston The protozoan flagellate parasite Giardia lamblia is the aetiologic agent for giardiasis, an infectious disease with significant burden in developing countries. Currently, clinical signs and symptoms accompanied by microscopy are used to diagnose giardiasis. However, microscopic stool analysis is limited by low sensitivity, labor intensive and high turn-around time. Moreover, polymerase chain reaction (PCR) has been applied in the detection of G.lamblia, but its cost and technical skills have compromised its clinical utilization. One-step rapid diagnostic tests (RDT) such as the SPANBIO™ have been developed for infection screening purposes due to high turn-around time and use in point-of-care testing. SPANBIO™ RDT is based on the principle of immuno-chromography. However, its utility in detection of giardiasis in Kenya has not been evaluated. This was cross-sectional health facility-based study that sought to evaluate the diagnostic performance of SPANBIOTM RDT in detection of Giardia lamblia with microscopy as the gold standard and PCR as a reference standard in a clinical setting. The specific objective were; To determine the specificity of SPANBIOTM RDT against microscopy and PCR in detecting G. lamblia in diarrheal patients, to determine the sensitivity of SPANBIOTM RDT against microscopy and PCR in detecting G. lamblia in diarrheal patients, to identify factors influencing SPANBIOTM RDT test performance in diarrheal patients. Data collection of one hundred and forty-seven stool specimens from microscopy confirmed G. lamblia infected =78 and uninfected =69 individuals, were documented including demographic and clinical information. The patients collected about 10gms of sample (after instructions) from which only 2gms (peas size) were processed and examined macroscopically and microscopically by the direct stool analysis procedure. Subsequently, the stools specimens were analyzed using the SPANBIO™ one-step RDT according to the manufacturer’s protocols. Total genomic DNA extraction was done on 2 gms of stool and PCR was performed by amplification of GDH (5′-TCAACGTCAACCGCTTCCT-3′) gene. Relative to the gold standard SPANBIOTM RDT illustrated a sensitivity of 66.7% (95% CI; 55.1-76.9%) and specificity of 98.6% (95% CI; 92.3-100%) with positive predictive value and negative predictive value of 98.1% (95% CI; 88.1%-99.7%) and 72.3% (95% CI; 65.6-78.1%) respectively; The test agreement between the SPANBIOTM and microscopy was high, and is indicated by Cohenʹs kappa coefficient = 0.6388; P<0.0001). When compared to PCR, RDT had sensitivity and specificity of 78.2 %, (95% CI; 67.4-86.8%) and 89.7% (95% CI; 80.2-95.8%) respectively and a positive predictive value and negative predictive value of 89.7 % (95% CI; 81.0-94.7%) and 78.5% (95% CI; 70.4-84.8%), respectively. The test agreement between the SPANBIOTM and PCR was high, and is indicated by Cohenʹs kappa coefficient = 0.6750; P<0.0001). Analyses to determine factors influencing SPANBIOTM RDT performance indicated that mucus, 2.982(95% CI; 0.089-0.440) and fecal pus 2.318(95% CI; 0.035-0.439) in stool affected its performance, G. lamblia was commonly found in semi-formed and loose stool. Therefore, there should also be a way of improving sample quality for giardiasis diagnosis. In conclusion the results obtained have shown that although mucus and fecal pus influence test performance, SPANBIOTM RDT is equally can be used in the diagnosis of G. lamblia. This study has significance as it has shown that there is an alternative technique that can produce timely, reliable and non-laborious results in the diagnosis of G. lamblia, which can be used in areas without microscopic capabilities. Masters Thesis

Xpert mtb/rif performance in detection of mycobacteriumtuberculosis in sputum pellets using a reduced sample reagent in smear negative samples in Kisumu, western Kenya

2022-12-20T14:39:06Z

Xpert mtb/rif performance in detection of mycobacteriumtuberculosis in sputum pellets using a reduced sample reagent in smear negative samples in Kisumu, western Kenya SITATI, N. Ruth Masters Thesis

Correlation of vitamin D levels and immune status in Hiv positive children aged 3 to 14 years attending Jaramogi Oginga Odinga teaching and referral hospital in Kisumu county, Kenya

2022-12-20T14:34:28Z

Correlation of vitamin D levels and immune status in Hiv positive children aged 3 to 14 years attending Jaramogi Oginga Odinga teaching and referral hospital in Kisumu county, Kenya SONGOREH, Maurice, .Asamuka Vitamin D deficiency is a worldwide phenomenon and it’s more prevalent in Human Immunodeficiency Virus (HIV) positive individuals. This prevalence could be higher in children from sub-Saharan Africa where the HIV burden is highest. In Kenya the occurrence of Vitamin D deficiency has been reported among children, diabetics and cancer patients. Children are the worst affected by HIV because the disease progresses faster due to their naïve immune systems. Alongside other factors this is compounded by deficiency diseases, most notably Vitamin D(25OHD) which acts as an immunomodulator of the adaptive immune system where it directly affects T cell activation and antigen presenting cells. Vitamin D deficiency has been associated with poor clinical outcome among adults living with HIV. However, it is not clearly known how this would play out in children. This was a comparative cross-sectional studywhere98HIV positive children aged 3 to 14 years attending the Jaramogi Oginga Teaching and Referral Hospital and unmatched98HIV negative children in the same age visiting the outpatient clinic at the same hospital were consecutively sampled and differences in the Vitamin D levels determined using blood samples. Correlation was then done between Vitamin D,CD4 and viral load for the HIV positive group. Vitamin D was determined using the ELISA technique, while CD4 levels was determined using 3-colour flow cytometry. HIV viral load levels were determined by real-time PCR. Clinical history was collected from the participants medical records. Independent samples T-test was used to compare Vitamin D means while Pearson correlation was used to correlate Vitamin D, CD4 and viral load. The HIV uninfected group had mean Vitamin D levels of 30.88 ng/ml (30.88±6.62 ng/ml) with deficiency (<20 ng/ml) and insufficiency (21-29 ng/ml) rates at 5.1% and 37.8% respectively. The HIV infected group had mean Vitamin D levels of 28.21 ng/ml (28.21±6.39 ng/ml) with deficiency and insufficiency rates at 13.3% and 46.9% respectively. There was a significant difference between the mean Vitamin D levels of the two groups(p=0.004). There was no correlation between Vitamin D and CD4count (r=.166, N=98, p=0.101), and Vitamin D and viral load (r=-.115, N=98, p=0.776). In conclusion prevalence of Vitamin D deficiency and insufficiency is higher in HIV infected children than in uninfected children and there is no correlation between Vitamin D status and immune status in HIV infected children. These findings suggest the assessment of Vitamin Din children as the adverse health effects extend beyond bone health. Assessment of Vitamin D in this demographic could help improve the overall health status of children especially the immunosuppressed. Masters Thesis

Influence of intermittent preventive treatment on transplacental transfer of anti-measles immunoglobulin-g antibodies in mother-infant pairs from Ahero sub-county hospital in Kisumu county, western Kenya

2022-05-12T06:56:30Z

Influence of intermittent preventive treatment on transplacental transfer of anti-measles immunoglobulin-g antibodies in mother-infant pairs from Ahero sub-county hospital in Kisumu county, western Kenya ARIERA, Bonface Ochieng Measles still remains one of the leading causes of vaccine preventable deaths globally with 140,000 deaths in 2019. These deaths occur despite the existence of a safe and effective measles vaccine. In Kenya, annual measles incidence ranges from 2-65 cases per million persons with about 76% of these being reported in unvaccinated infants aged less than a year. Primary protection against measles during infancy is mediated by maternally derived antibodies prior to vaccination with the levels in infants correlating with maternal antibody levels. The efficiency of transfer of measles specific antibodies is dependent on a number of factors including malaria infection. World Health Organization recommends malaria chemoprophylactic treatment in pregnant women in malaria endemic areas like western Kenya which may improve or suppress the vertical transfer of antibodies against other infectious diseases like measles. This study investigated the levels and vertical transfer of anti-measles Immunoglobulin-G to infant in expectant women taking either Dihydroartemisinin Piperaquine(DP) or Sulphadoxine Pyrimethamine (SP) for malaria Intermittent Preventive Treatment in pregnancy (IPTp). Specifically, this study compared the levels of anti-measles antibodies in mother-infant pairs undertaking IPTp-SP; compared the levels of anti-measles antibodies in mother-infant pairs undertaking IPTp-DP; compared the levels of anti-measles antibodies in mother-infant pairs between the IPTp-DP and SP treated participants, and finally investigated the influence of confounding variables (placental malaria, maternal age, hyper/po-gammaglobulinemia, parity and gestational age)- on the vertical transfer of anti-measles antibodies in women attending Ahero Sub-County Hospital in Kisumu County, Western Kenya. Using convenient sampling, samples from 132 mothers and 66 infants’ samples were included in this retrospective-prospective study design. The levels of antibodies against measles; EBV and malaria antibodies (internal positive controls) in maternal venous, cord (neonatal), and infants blood samples at one and six weeks were quantified using Luminex technology. Total IgG levels were determined in maternal venous blood by Enzyme-linked Immunosorbent Assay (ELISA). Wilcoxon paired test was used to compare the median levels of antibodies in mother-infant pairs to measles, EBV and malaria within the DP and SP treated groups. Using Wilcoxon paired test, the levels of MV (measles), EBV and malaria antibody levels were comparable between the mother-infant pairs in the SP treatment arm (P= 0.47, 0.97, 0.16, 0.47 and 0.73 for MV, AMA1, MSP1, EBNA1 and ZEBRA respectively). The levels were also comparable for the mother-infant pairs treated with DP (P= 0.71, 0.66, 0.96, 0.94, 0.71 for MV, AMA1, MSP1, EBNA1 and ZEBRA, respectively). Mann-Whitney U test comparing median antibody levels between the DP and SP treated mothers and their respective infants only showed a significant difference at enrollment (baseline) for MV and EBNA1 (P= 0.0057 and 0.0035, respectively). Anti-MV, AMA1, MSP1, ZEBRA and EBNA1 were however comparable at birth and among the infants. Spearman correlation analysis showed a positive correlation between maternal and neonatal antibody levels. Multivariate linear regression analysis showed no influence of the confounding variables investigated on transplacental transfer of antibodies. These findings suggest that malaria chemoprophylaxis with either DP or SP does not affect the levels and subsequent transfer of MV as well as EBV and malaria specific antibodies. This study also demonstrates that more than 20% of the infants by six weeks postpartum are highly susceptible to measles infection thereby pointing to the need for a booster dose of measles vaccine in women of child bearing age before conception to increase the duration of protection in infants before vaccination. In addition, this data confirms the safety of IPTP- DP/SP which makes them suitable for use in malaria endemic settings after considering their efficacy in malaria management.

Epstein bar virus strains in peripheral blood and saliva Of mother-child pairs in malaria holoend~mic regions With varying burkitt's lymphoma incidence rates

2022-04-25T09:23:44Z

Epstein bar virus strains in peripheral blood and saliva Of mother-child pairs in malaria holoend~mic regions With varying burkitt's lymphoma incidence rates NAMUYENGA, Toko Eunice Endemic Burkitt's lymphoma is a polymicrobial childhood cancer disease mainly associated with EBV and plasmodium falciparum infections whose combined effects profoundly affect the B cell compartment. The exact role of these microbes in Burkitt's lymphoma pathogenesis as well as other EBV associated malignancies is not well understood. Genetic polymorphisms have led to the identification of:EJ3V type. 1 and 2 which may be specifically associated with some virus positive tumors. EBV strain prevalence and the effect of malaria infections on EBV specific immune T' cell responses in children at risk for BL in malaria holoendemic areas have not been studied. This study determined the prevalence of EBV type 1 and type 2, malaria infection prevalence and interferon gamma T cell responses in children and their mothers from BL 'hot' and 'cold' spots. Since EBV can be detected in both saliva and blood, this study compared the differences in virus types between these compartments as well. In a cross sectional study design, samples were collected from 58 Burkitt's lymphoma cold spot area participants and 37 Burkitt's lymphoma hot spot area participants. DNA was extracted. from peripheral blood and saliva and PCR amplification was done on all samples. PCR primers were used to amplify a region of the EBNA3C gene that can distinguish between the two strains based on the base pairs of the PCR product. EBV type 1 and 2 were identified based on length differences within the EBNA3C gene. Malaria infections were also determined by blood smears and compared with EBV infections and specific T cell responses from ELISPOT assays. EBVDNA was detected in 94.6% of the hot spot area samples and 86.2% of the cold spot area samples. 75.15% of the participants hadEBV in blood compared to 66.1% in saliva. EBV type 1 .and 2 multiple infections were detected in 94.6% of the hot spot area participants compared to 51.7% of the cold spot area participants. Single type 1 and type 2 EBV infections were detected at low frequencies in the cold spot area. Comparison of the type ofEBV found in mothers and children showed only 61.5% matchin blood and saliva all being typel and 2 multiple infections. In the cold spot area, 38.1% had matching strains in saliva and 57.1 in blood. 37.5% had type 1 single infections, 25% type 2 single infections, and 37.5% type 1 and 2 multiple infections in saliva. On the other hand, in the hot spot samples, 8.3% had type 1 single infections, 8.3% had type 2 single infections and 83.3% type 1 and 2 multiple infections. Chi-square analysis at P<0.05 showed significantly high detection of EBV DNA in the hot spot area samples compared to the cold spot area samples.' Type 1 and 2 co-infections were predominant in the hot spot blood and saliva samples. A high percentage of children from both study sites were co-infected with both EBV type 1 and 2. A high proportion of . mother-child pairs had matching EBV types in both blood and saliva compared to the hot spot (P< 0.05). Malaria infection prevalence was relatively high in the hot spot area compared to the cold spot area. EBV specific T cell responses were reduced in the malaria infected individuals compared to the uninfected individuals. Thisstudy concludes . . that EBV type 1 and 2 co-infections are highly prevalent in the malaria holoendemic region with high BL incidence rates and there is uniform EBV type transmission from the mothers to their children. EBV type distribution was not however dependent on the prevalence of malaria. EBV specific T cell immune responses tent to be suppressed by malaria infections.