A Systematic Review of the Frequency of Regulatory T Cells in Hepatitis B and Hepatitis C

Background: Regulatory T cells (Tregs) play an important role in sustaining the hepatitis B and C viruses (HBV and HCV) persistence and protecting the liver tissues from cytokine-associated detrimental effects through unclear mechanisms. This paper aims to review the frequency of Tregs during the course of HBV and HCV infection.Method: Electronic databases were searched to identify studies investigated the frequency of intrahepatic and peripheral Tregs of the patients infected with HBV and/ or HCV.Results: The majority of studies reported the increase of intrahepatic and peripheral Tregs in acute and chronic infection of HBV and HCV. The decrease of peripheral Tregs occurred in patients with chronic hepatitis B who respond to interferon α or nucleos(t)ide analogues treatment as well as those with chronic hepatitis C who were treated with interferon, ribavirin or liver transplantation.Conclusion: Infection with HBV and HCV appears to induce the production of Tregs in blood and hepatocytes whereas treatment may decrease Tregs levels. As the optimum balance between regulatory and effector T during HBV and HCV infection is crucial for preventing liver damage, further studies should be directed on the development of Tregs during HBV and HCV infection as well as their involvement in immunomodulatory strategies for combating HBV and HCV

The Effect of Exclusive Breastfeeding on Wasting in Children Under Five: A Meta-Analysis Study

Background: Wasting is a form of malnutrition that has become a serious global health problem for the past few decades. Global prevalence of wasting is reported to reach 52 million or 7.7% of the under-fives. Global coverage of exclusive breastfeeding was still as low as 40%. The purpose of this study was to due a meta-analysis on the effect of exclusive breastfeeding on wasting in children under five.Subjects and Method: This was a meta-analysis study conducted from PubMed, SCOPUS, DOAJ, BASE, EBSCO, Emerald, ProQuest, ScienceDirect, ResearchGate, and Google Scholar databases. This study sought a systematic review with observational study design and multivariate-adjusted odds ratio (AOR) analysis. Wasting was defined as weight for height (WHZ) Z score <-2. The data were analyzed by RevMan 5 meta-analysis program with a fixed effect and random effect analysis model.Results: There were 13 articles included in this study. Based on 3 study design, exclusive breastfeeding could prevent the risk of wasting in children under five with each meta-analysis result from a cross-sectional study 0.42 times (OR = 0.42; 95% CI = 0.28 to 0.62; p <0.001), case-control studies 0.38 times (OR = 0.38; 95% CI = 0.27 to 0.54; p <0.001), and cohort studies 0.75 times (OR = 0.75; 95% CI = 0.55 to 1.04; p = 0.08). There was no publication bias in the results of the cross-sectional and case-control study meta-analysis, but there was in the results of the meta-analysis of the cohort study.Conclusion: Exclusive breastfeeding can prevent wasting in children under five.Keywords: exclusive breastfeeding, wasting, observational studyCorrespondence: Amallia Wijiwinarsih.Masters Program in Public Health, Universitas Sebelas Maret. Jl. Ir, Sutami 36A, Surakarta, Central Java 57126.Email:  [email protected]. Mobile: 0857261706.Journal of Maternal and Child Health (2019), 4(2): 87-96https://doi.org/10.26911/thejmch.2019.04.02.0

Giving Probiotic for a Better Therapy of Bacterial Vaginosis

Bacterial vaginosis (BV) is a common problem in women that is characterized by a decreased colonization of Lactobacillus in the vagina. Such condition may lead to various complications including endometritis, pelvic inflammatory disease, miscarriage, preterm birth, chorioamnionitis, premature rupture of the membrane, postpartum endometritis and increasing failure of in vitro fertilization. Treating BV using antimicrobials issomehow effective but often causes recurrences and antibiotic resistance. This paper examines the use of probiotic consisting of Lactobacillus spp. for the treatment of BV by conducting a systematic review of research articles published in online databases. After the review process, there are 18 articles with a total of 2,442 women as participants selected in this present report. There are six mechanisms involved in BV treatment using probiotic; i.e., (1) competition mechanism with pathogens, (2) stabilization of mucin layer, (3) production of antibacterial substances for killing and preventing the colonization of pathogenic bacteria, (4) reduction of the vaginal pH, (5) stimulation of the immune system, and (6) restoration of vaginal homeostasis. Previous studies were evaluated and the findings suggest that probiotics are better than antimicrobials in treating BV due to the absence of reported side effects. In addition, giving probiotics especially Lactobacillus sp. can be a better therapy of BV

Relative Efficacy of Probiotics Compared with Oral Rehydration Solution for Diarrhea Treatment in Children under Five Years Old: a Meta-Analysis from Developing Countries

Background: Children under five experience an average of three episodes of diarrhea each year in developing countries. Based on World Health Organization (WHO) guidelines, therapy for acute diarrhea is oral rehydration fluid and zinc to treat fluid and electrolyte loss. But unfortunately, this therapy does not reduce the duration of diarrhea without the intestinal barrier function of pathogenic microorganisms. The ability to inhibit pathogens is one of the three main mechanisms of probiotics. This study aims to examine the efficacy of using probiotics compared to oral rehydration solution (ORS) in the treatment of acute diarrhea in children under five in developing countries.Subjects and Method: This study is a meta-analysis conducted using PRISMA systematic guidelines. The process of searching for articles was carried out between 2009 and 2019 using a database search engine consisting of PubMed, British Medical Journal (BMJ), CAB Direct, Oxford Academy, Clinical Key, ScienceDirect, and Scopus. Based on a database search, six articles that meet the criteria of the Randomized Controlled Trial (RCT) and research conducted in developing countries were found. The study involved 1234 children who were divided into two groups: 762 probiotic groups and 472 ORS groups. The analysis was performed with the Review Manager (RevMan) software 5.3. The results were assessed using Standardized Mean Difference (SMD).Results: There was heterogeneity between experiments (I2 = 91%; p <0.001) so Random Effects Model (REM) was used. Probiotics could reduce the duration of acute diarrhea in infants rather than just ORS, with a pooled estimate of 1.13 (SMD = -1.13; 95% CI = -1.54 to -0.72; p <0.001).Conclusion: Probiotics can reduce the duration of acute diarrhea in infants in developing countries rather than just the administration of ORS.Keywords: Diarrhea, probiotics, oral rehydration solution, meta-analysis.Correspondence:Nurul Aini Suria Saputri. Masters Program in Public Health, Universitas Sebelas Maret. Jl. Ir. Sutami 36 A, Surakarta 57126, Central Java. Email: [email protected]. Mobile: 085­743401971Indonesian Journal of Medicine (2019), 4(4): 354-363https://doi.org/10.26911/theijmed.2019.04.04.08

Evaluasi MetodeGeneXpert MTB/RIF dengan SampelRaw Sputum untuk Mendeteksi Tuberkulosis Paru

Tuberkulosis (TB) merupakan penyakit menular yang masih menjadi masalah utama di Indonesia dan dunia. Tantangan utama dalam mendiagnosis TB secara konvensional yaitu rendahnya sensitivitas deteksi pada pemeriksaan mikroskopis dan lamanya waktu yang diperlukan untuk kultur. Penelitian ini bertujuan untuk mengevaluasi metode GeneXpert MTB/RIF untuk mendeteksi Mycobacterium tuberculosis dengan sampel dahak langsung di RSUD Dr. Moewardi (RSDM), Surakarta. Analisis observasional dengan pendekatan kohort retrospektif menggunakan data sekunder hasil pemeriksaan GeneXpert MTB/RIF pada sampel dahak langsung di Laboratorium Mikrobiologi Klinik RSDM pada tahun 2012-2015. Sampel didapatkan dari pasien yang memenuhi kriteria suspek multidrug-resistant tuberculosis (MDR-TB) dan sampel tersebut telah dikultur di Balai Laboratorium Kesehatan (BLK), Jawa Tengah. Data dianalisis dengan OpenEpi versi 3, Epi Info 7, dan MedCalc. Pada penelitian ini didapatkan sensitivitas, spesifisitas, nilai duga positif dan negatif (NDP dan NDN) serta akurasi metode GeneXpert MTB/RIF sebesar 93,62%, 27,17%, 68,89%, 71,21%, dan 69,21%. Prevalensi TB paru pada sampel yang diperiksa sebesar 63,27%. Rendahnya spesifisitas GeneXpert MTB/RIF mengindikasikan perlunya kultur sebagai baku emas. Namun demikian, perlu standarisasi pemrosesan sampel dahak dalam segi teknik dan waktu pengambilan sampel disertai data klinis yang memadai untuk melihat riwayat terapi yang telah diberikan pada pasien

The role of genital tract infection inpreterm delivery: aretrospective study

 Preterm delivery is associated with higher mortality and morbidity of neonates, also increasestheir risk of having growth and development impairment.This study aimed toidentify the role ofgenital tract infection in preterm delivery. A retrospective study was conducted on medical records of mothers who had preterm delivery in a tertiary hospital in Surakarta, Indonesia during 2017. The data collected were mothers’ age and their gestational age, the history of current pregnancy, the number of previous abortion(s), mothers’ body temperature, the extent of abnormal vaginal discharge, and laboratory findings (white blood cell count, platelet count, red blood cell count, hemoglobin count, hematocrit level, urinalysis and microbiology results). The statistical differences amongst categorical and numerical data were analyzed using the Chi-Square test and the Mann-Whitney test. Based on the patient’s history and the examination results, we suspected genital tract infections in22.52% (25/111)of subjects. All of them had abnormal vaginal discharge despite only one case had been confirmed as streptococcal infection. This study found that the majority of mothers with probable genital tract infection hadpreterm premature rupture of the membrane whilst preeclampsiawas more evident in those without genital tract infection. We conclude that genital tract infection during pregnancy is a significant contributor to the occurence of premature birth so that microbiological testing is needed to confirm the diagnosis

Characteristics of patients with Hepatitis B and C at Dr. Moewardi General Hospital in Surakarta, Indonesia

Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections remain a significant health burden in the world, which is mainly attributed to patients who develop chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). The epidemiology of hepatitis B and C in Surakarta, Central Java Province, Indonesia has never been reported. This study aimed to investigate the demographic, clinical, and laboratory characteristics of patients with hepatitis B and C who were admitted to Dr. Moewardi General Hospital, Surakarta in 2019. The medical records of patients with hepatitis B (n=94) and hepatitis C (n=75) were examined, and the data were analyzed using the chi-square and Mann-Whitney tests. The patients with hepatitis C were generally older, more likely to develop jaundice and ascites, and had higher levels of serum urea, creatinine, AST, and total bilirubin compared to those with hepatitis B. In conclusion, patients with HCV infection had worse clinical presentation and laboratory profiles than those with HBV infection. However, further research is needed on a wider scale to confirm this result

PEMBERDAYAAN YAYASAN SEBAGAI PELOPOR GERAKAN MASYARAKAT HIDUP SEHAT DALAM UPAYA PENCEGAHAN PENYAKIT HIPERTENSI

ABSTRAKPenyakit Tidak Menular (PTM) merupakan penyebab kematian tertinggi di dunia, termasuk di Indonesia. Salah satu PTM yang cukup tinggi prevalensinya adalah hipertensi. Kabupaten Karanganyar merupakan salah satu kabupaten di provinsi Jawa Tengah dengan jumlah kasus hipertensi yang cukup tinggi dan prevalensinya terus meningkat dari tahun ke tahun. Pemerintah Indonesia dan pemerintah kabupaten setempat telah mencanangkan Gerakan Masyarakat Hidup Sehat (Germas) sebagai upaya pengendalian PTM. Yayasan Usaha Umat Karanganyar (YUUK), sebuah organisasi masyarakat yang berlokasi di Tasikmadu, Karanganyar, telah memiliki program kegiatan pendidikan dan pelatihan yang diperlukan oleh masyarakat setempat, namun program pendidikan dan pelatihan tersebut belum menjamah aspek kesehatan masyarakat. Untuk mengoptimalkan kesehatan masyarakat setempat melalui pemberdayaan organisasi kemasyarakatan, tim pengabdian masyarakat yang tergabung dalam grup riset Human Proteomics Fakultas Kedokteran Universitas Sebelas Maret (FK UNS) melakukan kegiatan penyuluhan dan konsultasi tentang hipertensi dan serta pelatihan penggunaan tensimeter digital dan oximeter. Kegiatan tersebut bertujuan untuk meningkatkan peran serta YUUK dalam pencegahan dan deteksi dini penyakit hipertensi. Sasaran kegiatan ini adalah para remaja di sekitar masjid Al Walidah Tasikmadu Karanganyar. Hasil dari kegiatan tersebut adalah meningkatnya pemahaman peserta mengenai hipertensi dan keterampilan memantau kesehatan kardiovaskuler melalui penggunaan tensimeter digital. Evaluasi kegiatan ini dilakukan dengan cara menghubungi pengurus YUUK untuk mendapatkan laporan tentang implementasi pemanfaatan 2 unit tensimeter digital dan 1 unit oximeter yang diberikan oleh tim pengabdi kepada pengelola Yayasan untuk dimanfaatkan sebaik-baiknya bagi masyarakat sekitar. Tim pengabdi juga terus memonitor prevalensi hipertensi dari tahun ke tahun di Kabupaten Karanganyar. Kata kunci: hipertensi; pemberdayaan masyarakat; penyakit tidak menular ABSTRACT Non-communicable diseases (NCD) are the major causes of mortality in the world, including in Indonesia. One of the most prevalent type of NCD is hypertension. Karanganyar is a district in Central Java province with high incidence of hypertension and its prevalence continues to rise over years. Both central and local governments have established "Gerakan Masyarakat Hidup Sehat (Germas)", a community-based movement to promote healthy lifestyle, to control NCD. "Yayasan Usaha Umat Karanganyar (YUUK)", a community organization located in Tasikmadu, Karanganyar, provides education and training for locals but their main focus has not reached health programs. To optimize the locals' health through empowerment of community organizations, we conducted education and counseling about hypertension as well as training on using digital sphygmomanometer and oxymeter. The activities aimed to increase YUUK's roles in the prevention and early detection of hypertension. The activities involved teenagers living in the catchment area of Al Walidah mosque, Tasikmadu, Karanganyar. The activites resulted in an increased understanding about hypertension and skills to monitor cardiovascular health by using a digital sphygmomanometer,  Evaluation was performed by contacting the YUUK manager who is responsible for reporting the use of 2 units of digital sphygmomanometers and 1 unit of oxymeter that were given during the occasion so that the equipment can be used by locals. We also continuosly monitor the annual prevalence of hypertension in Karanganyar district. Keywords:  hypertension; community empowerment; non-communicable disease

Evaluation of next-generation sequencing technology in determining infectious causes of fever

Undifferentiated fever (UDF) is a common complaint in clinical practice, but its aetiology is not always determined due to non-specific symptoms and laboratory findings. While fever of unknown origin (FUO) is a common medical term for fever without obvious cause, this condition is distinguished from acute undifferentiated fever (AUF) in terms of duration, progression of illness and underlying causes. In FUO, fever must exist for more than 3 weeks and can persist for a very long period unless the underlying cause is found and eliminated. In contrast, AUF is more limited in duration and many episodes spontaneously resolve, presumably due to self-limiting infectious diseases. The problem of determining the infectious causes of fever has received considerable attention, particularly in tropical countries. Previous studies in South and Southeast Asia reported high prevalence of infection as the main cause of AUF. This prompted the hypotheses that infection-related AUFs are common in the tropical region of Far North Queensland, Australia, and that a significant proportion of AUFs in this region are undiagnosed. Diagnosing infectious causes of fever is a challenge for clinicians. With hundreds of possible agents and a limited number of specific tests that can be performed, it is very likely that doctors will miss the true cause of fever. Moreover, current diagnostic approaches rely on prior knowledge of the pathogens being sought, thus precluding the detection of unsought or novel pathogens. Thus, the prevalence of undiagnosed undifferentiated fever (UUDF) indicates either that clinicians are failing to order appropriate tests, that current diagnostic methods are inadequate, or that there are causes of fever that are yet to be discovered. This diagnostic challenge necessitates good clinical skills, knowledge of the pattern of signs and symptoms associated with particular infections and a broad diagnostic tool for determining infectious causes of fever. Since there is a wide range of pathogens, the diagnostic tool should be able to distinguish one pathogen from another as well as identify multiple pathogens simultaneously. This can be achieved if the diagnostic tool can 'read' unique characteristics of pathogen(s) present in the sample as distinguishable nucleic acid sequences so it can facilitate the identification of organism(s). Next-generation sequencing (NGS) technology has the capability to produce large amounts of nucleic acid sequences in a relatively short time. This technology has been applied to the study of biological diversity in environmental samples. The extent of diagnostic problems and the capability of NGS technology to detect the presence of nucleic acids in any environment have brought the theory into clinical application. The potential of NGS as a broad-scale diagnostic tool prompted the hypothesis that NGS is a practical method for investigating pathogens causing fever. There are several NGS platforms available, but little is known about their effectiveness and efficiency with regards to pathogen detection in clinical samples. The primary aim of this study was to assess the capacity, sensitivity, and more importantly, the specificity of the Illumina HiSeq platform for broad-scale characterisation of pathogens associated with fever. Secondary aims included: to describe the epidemiology of AUF and UUDF in Far North Queensland, Australia, and to optimise the preparation procedure for samples that will be subjected to NGS assay. The study was conducted in three stages, comprising two preliminary studies and the main study. The first preliminary study was a retrospective study of fever patients presenting to Cairns Hospital over the three-year period between 1 July 2008 and 30 June 2011. The findings suggest that AUF is common in the population of Far North Queensland, Australia. A robust definition of UUDF is proposed based on the clinical and laboratory characteristics of patients reviewed in this study, including the following criteria: 1) a fever of ≥ 38.0°C or symptoms suggestive of fever; 2) a duration of fever of ≤ 21 days; 3) a failure to reach a diagnosis after performing clinical evaluation and laboratory investigations, including complete blood count, serum biochemistry, urinalysis, blood culture, chest X-ray; 4) a request by the clinician of specific test for at least one infectious agent and; 5) a failure to make a specific diagnosis. The proportion of UUDF was 56.8% (193/340), indicating the need for a broad diagnostic tool to determine infectious causes of AUF. In general, the findings provide valuable information regarding the feasibility of conducting a fever study using NGS technology at Cairns Hospital. The second preliminary study was conducted to determine the most suitable type of blood specimen for NGS analysis. It was anticipated that there would be small quantities of pathogen nucleic acids present among abundant human nucleic acids background. Therefore, it was important to minimise the quantity of human nucleic acids in order to increase the sensitivity of detection of the pathogen. Six healthy volunteers participated in this second preliminary study, which aimed to measure levels of double-stranded DNA in plasma and serum. Specimens were taken using different methods of blood collection: with a syringe and with a vacuum system, and with and without applying a tourniquet. DNA concentration in the samples was measured using microplate fluorescence assays using SYBR Green I as the fluorescent dye. This study found that DNA concentration in plasma was significantly lower than that in serum (p < 0.05). However, the method of blood collection did not significantly affect DNA concentration. A main component of this thesis was a prospective study involving the use of a NGS platform to determine infectious causes of AUFs. Isolation of DNA and RNA from plasma/serum samples was performed using QIAamp® DNA Mini Kit (Qiagen) and TRIzol® LS reagent (Life Technologies) respectively. Following nucleic acid extraction, amplification of DNA was conducted according to the SeqPlex Enhanced DNA Amplification Kit (SEQXE) protocol (Sigma-Aldrich). DNase treatment, cDNA synthesis and amplification were performed on RNA samples according to SeqPlex RNA Amplification Kit (SEQR) protocol (Sigma-Aldrich). Sequencing was conducted on 22 DNA/cDNA samples that met the standard input determined by the sequencing company. These samples originated from 17 patients, comprising seven positive control samples from patients who had specific diagnoses and 10 samples from patients for whom diagnoses had not been achieved. Data analysis was conducted using the Kraken program and the traditional assembly-alignment pipeline. The study findings demonstrate the limitation and utility of NGS technology in determining the aetiology of AUF. Various viruses and bacteria were found in every sample, so considered selections were made on pathogens for which there were supporting reads consistent with clinical data and pathology findings. Aetiological diagnosis was verified in 85.7% (6/7) of controls. Among the undiagnosed participants, deep sequencing identified some plausible causes of fever in 60% (6/10) of subjects, including Escherichia coli bacteraemia and scrub typus that eluded conventional tests. Further, the NGS technology generated valuable information for studying microbial diversity in human blood. Further work could be directed at optimising sample preparation and improving sequencing efficiency, as well as developing efficient bioinformatics tools for analysing sequence data. It is hoped that NGS technology can be adopted in clinical practice at more affordable costs and with timely delivery of results

Evaluation of next-generation sequencing technology in determining infectious causes of fever

Undifferentiated fever (UDF) is a common complaint in clinical practice, but its aetiology is not always determined due to non-specific symptoms and laboratory findings. While fever of unknown origin (FUO) is a common medical term for fever without obvious cause, this condition is distinguished from acute undifferentiated fever (AUF) in terms of duration, progression of illness and underlying causes. In FUO, fever must exist for more than 3 weeks and can persist for a very long period unless the underlying cause is found and eliminated. In contrast, AUF is more limited in duration and many episodes spontaneously resolve, presumably due to self-limiting infectious diseases. The problem of determining the infectious causes of fever has received considerable attention, particularly in tropical countries. Previous studies in South and Southeast Asia reported high prevalence of infection as the main cause of AUF. This prompted the hypotheses that infection-related AUFs are common in the tropical region of Far North Queensland, Australia, and that a significant proportion of AUFs in this region are undiagnosed. Diagnosing infectious causes of fever is a challenge for clinicians. With hundreds of possible agents and a limited number of specific tests that can be performed, it is very likely that doctors will miss the true cause of fever. Moreover, current diagnostic approaches rely on prior knowledge of the pathogens being sought, thus precluding the detection of unsought or novel pathogens. Thus, the prevalence of undiagnosed undifferentiated fever (UUDF) indicates either that clinicians are failing to order appropriate tests, that current diagnostic methods are inadequate, or that there are causes of fever that are yet to be discovered. This diagnostic challenge necessitates good clinical skills, knowledge of the pattern of signs and symptoms associated with particular infections and a broad diagnostic tool for determining infectious causes of fever. Since there is a wide range of pathogens, the diagnostic tool should be able to distinguish one pathogen from another as well as identify multiple pathogens simultaneously. This can be achieved if the diagnostic tool can 'read' unique characteristics of pathogen(s) present in the sample as distinguishable nucleic acid sequences so it can facilitate the identification of organism(s). Next-generation sequencing (NGS) technology has the capability to produce large amounts of nucleic acid sequences in a relatively short time. This technology has been applied to the study of biological diversity in environmental samples. The extent of diagnostic problems and the capability of NGS technology to detect the presence of nucleic acids in any environment have brought the theory into clinical application. The potential of NGS as a broad-scale diagnostic tool prompted the hypothesis that NGS is a practical method for investigating pathogens causing fever. There are several NGS platforms available, but little is known about their effectiveness and efficiency with regards to pathogen detection in clinical samples. The primary aim of this study was to assess the capacity, sensitivity, and more importantly, the specificity of the Illumina HiSeq platform for broad-scale characterisation of pathogens associated with fever. Secondary aims included: to describe the epidemiology of AUF and UUDF in Far North Queensland, Australia, and to optimise the preparation procedure for samples that will be subjected to NGS assay. The study was conducted in three stages, comprising two preliminary studies and the main study. The first preliminary study was a retrospective study of fever patients presenting to Cairns Hospital over the three-year period between 1 July 2008 and 30 June 2011. The findings suggest that AUF is common in the population of Far North Queensland, Australia. A robust definition of UUDF is proposed based on the clinical and laboratory characteristics of patients reviewed in this study, including the following criteria: 1) a fever of ≥ 38.0°C or symptoms suggestive of fever; 2) a duration of fever of ≤ 21 days; 3) a failure to reach a diagnosis after performing clinical evaluation and laboratory investigations, including complete blood count, serum biochemistry, urinalysis, blood culture, chest X-ray; 4) a request by the clinician of specific test for at least one infectious agent and; 5) a failure to make a specific diagnosis. The proportion of UUDF was 56.8% (193/340), indicating the need for a broad diagnostic tool to determine infectious causes of AUF. In general, the findings provide valuable information regarding the feasibility of conducting a fever study using NGS technology at Cairns Hospital. The second preliminary study was conducted to determine the most suitable type of blood specimen for NGS analysis. It was anticipated that there would be small quantities of pathogen nucleic acids present among abundant human nucleic acids background. Therefore, it was important to minimise the quantity of human nucleic acids in order to increase the sensitivity of detection of the pathogen. Six healthy volunteers participated in this second preliminary study, which aimed to measure levels of double-stranded DNA in plasma and serum. Specimens were taken using different methods of blood collection: with a syringe and with a vacuum system, and with and without applying a tourniquet. DNA concentration in the samples was measured using microplate fluorescence assays using SYBR Green I as the fluorescent dye. This study found that DNA concentration in plasma was significantly lower than that in serum (p < 0.05). However, the method of blood collection did not significantly affect DNA concentration. A main component of this thesis was a prospective study involving the use of a NGS platform to determine infectious causes of AUFs. Isolation of DNA and RNA from plasma/serum samples was performed using QIAamp® DNA Mini Kit (Qiagen) and TRIzol® LS reagent (Life Technologies) respectively. Following nucleic acid extraction, amplification of DNA was conducted according to the SeqPlex Enhanced DNA Amplification Kit (SEQXE) protocol (Sigma-Aldrich). DNase treatment, cDNA synthesis and amplification were performed on RNA samples according to SeqPlex RNA Amplification Kit (SEQR) protocol (Sigma-Aldrich). Sequencing was conducted on 22 DNA/cDNA samples that met the standard input determined by the sequencing company. These samples originated from 17 patients, comprising seven positive control samples from patients who had specific diagnoses and 10 samples from patients for whom diagnoses had not been achieved. Data analysis was conducted using the Kraken program and the traditional assembly-alignment pipeline. The study findings demonstrate the limitation and utility of NGS technology in determining the aetiology of AUF. Various viruses and bacteria were found in every sample, so considered selections were made on pathogens for which there were supporting reads consistent with clinical data and pathology findings. Aetiological diagnosis was verified in 85.7% (6/7) of controls. Among the undiagnosed participants, deep sequencing identified some plausible causes of fever in 60% (6/10) of subjects, including Escherichia coli bacteraemia and scrub typus that eluded conventional tests. Further, the NGS technology generated valuable information for studying microbial diversity in human blood. Further work could be directed at optimising sample preparation and improving sequencing efficiency, as well as developing efficient bioinformatics tools for analysing sequence data. It is hoped that NGS technology can be adopted in clinical practice at more affordable costs and with timely delivery of results