Diagnosis of malaria and typhoid fevers using basic tools: a comparative analysis of a retrospective data with a prospective evaluation in an endemic setting

Malaria and typhoid fever are among important and endemic diseases in the tropical countries such as Nigeria. Diagnosis of most cases of malaria/typhoid co-infections are based on clinical suspicion alone or unreliable diagnostic tools leading to poor or misdiagnosis. A retrospective analysis was conducted on the positivity rate for malaria parasite and typhoid fever among patients who attended the Bayero University, Kano (BUK) Health Clinic from January to December 2013. A prospective study was also carried out on 200 febrile patients and 80 apparently healthy subjects (controls) with the view to determine the diagnostic profiles of malaria and /or typhoid fever co-infection using standard protocols (microscopy, rapid diagnostic test (RDT) for malaria parasite (MP), Widal test, stool and blood cultures for typhoid fever). Both test and control subjects were evaluated for the presence or otherwise of the malaria and /or typhoid aetiologic agents. Results obtained from the retrospective study for the period January to December 2013 indicated that, of the 2362 tests conducted, 318 (13.5%) were positive for MP using RDT and 722 (30.6%) were positive for typhoid fever using Widal test. Co-infection rate obtained was 89 (3.8%). Of the 200 subjects evaluated for the prospective study however, 42 (21%) and 34 (17%) were positive for MP using microscopic technique and RDT respectively. Microscopy was established to be more sensitive than RDT. Eighty nine (45%) were Widal positive. The culture method for typhoid fever diagnosis was negative. Co-infection rate stood at 17 (8.5%). No association however was found between the two disease conditions (p=0.6032). Significant proportion of the patients 86 (43%) were neither positive for malaria nor for typhoid fevers. Three isolates of non-typhoid salmonella specie and three other bacterial isolates were recovered from the stool and blood samples of the patients respectively. Baseline titre value for Widal test at BUK community was established as 160 and above for O antibodies to S. typhi, hence the criterion employed in the prospective evaluation. Laboratory evidence of malaria and typhoid co-infection rate at a titre of ≥160 was 8.5% using microscopy and 7% using RDT. The use of RDT is simple and easy, but has less sensitivity, thus may leave some patients with malaria un-detected. Based on the results of these findings, vis a vis the proportion of individuals negative for both malaria and typhoid fevers, clinicians should revisit causes of febrile illnesses other than malaria or typhoid and hence the need to include other tests for the detection of other causes.Keywords: Malaria diagnosis, Typhoid fever, RDT, Widal test, Co-infections, Nigeri

Applications of Molecular Diagnostic Techniques for Infectious Diseases

Diagnosis is concerned with identifying the cause of a disease or precise and consistent outcomes that are results of direct or indirect actions, reactions and interactions between the cause of a disease and the host. That outcome, if accurate, would help the clinician in disease management, or the epidemiologist in identifying trends of diseases or the administrator in policy and decision making. Traditionally, infectious disease diagnosis involves identifying the causative agents of infectious diseases through the direct examination, culture and often immunological tests on clinical specimens. The traditional diagnostic  techniques have varied sensitivities and specificities which influence their choice and applicability in a particular setting for the diagnosis of infectious diseases. However, the limitations of many traditional techniques particularly low specificity and long turnaround time often necessitate initiation of treatment before results are made available. Molecular diagnostic techniques involve a variety of techniques that explore the use of nucleic acid molecules for the identification of a particular pathogenic organism. These techniques include nucleic acid-based typing system, nucleic acid analysis without amplification, polymerase chain reaction (PCR) and other nucleic acid amplification techniques. Applications of molecular detection methods for infectious diseases have resolved many of the problems of the traditional diagnostic techniques, due to their exquisite sensitivity and specificity that allow the accurate and timely detection of very small numbers of organisms. This paper examines the principles and applications of molecular biology techniques in the identification of the causative agents of  infectious  diseases either in a routine setting or as research tools.Keywords: Infectious diseases, Molecular diagnosis, Polymerase Chain Reactio

Malaria and Hepatitis B co-infection in patients with febrile illnesses attending general outpatient unit of the Murtala Muhammed Specialist Hospital, Kano, Northwest Nigeria

Malaria and Hepatitis B Virus (HBV) infections are co-endemic throughout much of the tropical and sub-Saharan Africa and both present major threat to public health. A study on the prevalence of HBV and Malaria co-infection was carried out on 200 patients presenting with fever at the General Outpatient Department (GOPD) of the Murtala Muhammed Specialist Hospital (MMSH), Kano using Gold Standard microscopy and rapid diagnostic test (RDT). The effect of mono and co-infection on hematological parameters was also investigated. Fifty one (25.5%) out of the 200 patients studied were Malaria  positive. Females had higher prevalence rate(18%) of Malaria infection than males with 7.5%. Age group 15-24 had the highest Malaria prevalence (11%) followed by age group 25-34 with 6.5%. Higher mean  parasite density (1,200/ìl) was recorded among subjects with monoinfection of Malaria than mean parasite density (518/ìl) obtained among the co-infected. Mean parasite density was higher in female than male subjects. Thirteen (6.5%) subjects were HBV positive. Males had higher rate of infection with 4.5% prevalence than females with 2.0%. Nine individuals representing 4.5% of the total population had  co-infection with higher prevalence (3%) among the males. Age groups 25-34 were observed to have high co-infection rate of 1.5% and the least prevalence was observed among the age group 15-24 with 0.5%  prevalence for both males and females. Hematological evaluation carried out on all the categories of  subjects shows significant difference in mean values of PCV (P=0.041), Hb (P=0.018) between the  co-infection group and those with malaria infection and control groups. However, no significant difference (P>0.05) was observed in the values of WBC, PLT and Red cell indices among the co-infected and other test group. It was concluded that co-infection with the two ailments had no profound effect on hematologic parameters.Keywords: Co-Infection, Hepatitis B, Kano, Malaria, MMSH, Prevalenc

Comparing Haemozoin count and Parasitaemia in the Prognosis of severe Plasmodium Falciparum Malaria in Children and non-immune adults in Kano-Nigeria

This study compared the value of pigment containing leucocytes (neutrophils and monocytes) counts and the malaria parasite density in the prognosis of severe malaria on four hundred and twenty (420) patients with clinical evidence of severe malaria in Kano-Nigeria. Three clinical groups comprising patients with impaired consciousness, patients with cerebral malaria and those with severe anaemia were identified. Samples were analysed for Malaria parasitemia and pigment count Giemsa’s thick and Leishman’s thin film respectively. Patients with impaired consciousness (n=217) recorded the highest malaria pigment count of 342.86 (±177.34) monocytes pigments/microliter. The highest parasite count of 234,962 (±264.5) per microliter was recorded among the cerebral malaria group. Patients with severe malaria and anaemia had the least neutrophils pigment and parasite counts of 219.0 (±140.96)/microliter and 212,232(±12.61)/microliter respectively. A linear relationship between the malaria parasite count and the Intraneutrophilic malaria pigment count in severe malaria was demonstrated. Pigment count proved a higher prognostic value in severe malaria compared to Parasitaemia.Keywords: Haemozoin, Plasmodium falciparum, Parasitaemia, Malari

Prevalence and Risk Factors of High Risk Human Papillomavirus Infections among Women Attending Gynaecology Clinics in Kano, Northern Nigeria

Cervical cancer is the most common female cancer in northern Nigeria, yet the pattern of infection with human papillomavirus, the principal aetiologic agent is unknown. This was a preliminary study conducted in two referral hospitals in order to establish base-line data on the prevalence and risk factors for the infection in Kano state, Nigeria. Fifty (50) randomly selected women aged 18 years and above were recruited from gynaecology clinics in Murtala Muhammad Specialist Hospital and Aminu Kano teaching Hospital. Relevant sexual and socio-demographic information were obtained from each subject using a questionnaire. Exfoliated cervical cells were harvested and processed using Polymerase Chain Reaction to identify the DNAs of high-risk HPV types 16 and 18. The prevalence rate of HPV infection was 76% [(38/50) at 95% CI=61.8-86.9] with 60.5% (23/38) having co-infections with both HPV type 16 and 18. Risk factors of the infection include low literacy level; living in rural settlements; low parity; early menarche (<15 years of age); early onset of first sexual intercourse (≤16 years of age) and multiple sexual partners. There was however, no statistically significant association between oral contraceptive usage and acquisition of the infections. Findings of this study suggest a high prevalence of HPV types 16 and 18 among women attending gynaecology clinic in Kano and thus called for more elaborate community based study in order to establish the magnitude of the problem from wider perspective.Keywords: Human Papillomavirus; Prevalence; Risk Factors, Northern Nigeri

District level external quality assurance (EQA) of malaria microscopy in Pakistan: pilot implementation and feasibility

Background Prompt, quality assured laboratory diagnosis is key to effective malaria case management and control, especially since the introduction of the more expensive artemisinin combination therapy (ACT). The malaria programme and its non-government partners, on the basis of WHO recommended Lot Quality Assurance methods, have developed a district level external quality assurance (EQA) system. This study was designed to assess the feasibility, under programme conditions, of an integrated district level external quality assurance and supervision approach for malaria microscopy. Design and Methods A prospective study conducted over seven months period (May-November 2007). In addition to the standard WHO EQA elements, three operational innovations were introduced, with the a district laboratory supervisor: a) onsite re-checking of slides, b) in ensuring uninterrupted availability of laboratory reagents and supplies at diagnostic centers, and c) supervision of administrative and technical components. The quantitative data for the study came from the service records/documents, whereas the qualitative data came from the key informant interviews. Results During the seven month period in four districts, a total of 8,118 slides were examined of which 209 (2.6%) were found positive for malaria parasites (slide positivity range between1.6% to 6.0%). The District Laboratory Supervisors in four districts reexamined a total of 1,770 slides (22%). The proportion of slides found discordant ranged from 0.5% to 1%. The quality of smear preparation was found acceptable in 73% slides. Conclusions A district-based EQA, based on lot quality assurance methods was implemented, using context-specific operational guidelines, tools and training modules, and other inputs from the malaria control programme and partners. This EQA and supervision approach was found to be feasible and acceptable to those involved. Further study is required on the microscopy quality and cost-effectiveness of adding external quality assurance and supervision to district malaria microscopy services

Impact of introducing integrated quality assessment for tuberculosis and malaria microscopy in Kano, Nigeria

Background: The World Health Organization (WHO) has recommended the integration of malaria microscopy quality assessment (QA) with that of other microscopically diagnosed diseases, but there is no evidence that it has been attempted. We assessed the feasibility of linking malaria microscopy into the existing tuberculosis (TB) microscopy QA system in Kano, Nigeria. Methodology: Five TB microscopy centres were selected for implementing the integrated TB and malaria microscopy QA scheme in the state. A model system was designed for selecting and blinded rechecking of TB and malaria slides from these laboratories. Supervision and evaluation was conducted at 3-month intervals for 24 months. Results: TB microscopy QA was strengthened in four laboratories. Full integration of the QA for TB and malaria microscopy was achieved in two laboratories, and partial integration in two other laboratories. The programme resulted in an increase in the specificity of both TB and malaria microscopy results. At the final assessment, 100% specificity was achieved for TB microscopy results in four laboratories. There was an increased concordance rate and decreased false positivity and false negativity rates of TB microscopy results in all five laboratories. Conclusions: It is feasible to integrate the QA system and training for TB and malaria microscopy for assessing and improving quality of both services. However, the integrated system needs testing in different settings in order to be able to develop sound recommendations to guide the complex scaling-up process

Correlation of nosocomial infection with prolonged hospital stay in Kano Nigeria

Nosocomial infections or Health Care prolonged hospital stay and has been implicated in increase in socio-economic disturbance, long term disability, and increased mortality rate. There is paucity information on the burden of HCAIs because of poorly developed surveillance systems and inexistent control methods. We aimed to investigate the prevalence of nosocomial infection due to prolonged hospital stay in selected tertiary hospitals of Kano metropolis. Retrospective data  were collected from three hospitals with a total number of admitted patients and the number of prolonged hospital stays during the month of study. A total of 401 clinical samples collected from patients admitted for ≥14 days and the age of ≥18 years from all study sites. Examples include wound swabs, urine samples, urine catheters, and nasal intubation. All the samples were processed by the standard bacteriological laboratory procedure of the Clinical laboratory standard institute. The results showed that the percentage of prolonged hospital stay in Kano 40.34%, Murtala Muhammad Specialist Hospital (MMSH) 50.54% with the least at Muhammad Abdullahi Wase Specialist (MAWSH) 28.91%. Age served as significant factors for acquired HCAIs; participants aged were 41- 70 years have a higher prevalence of nosocomial infections. From 138 positive isolates observed, Murtala Muhammad Specialist Hospital (MMSH) has height nosocomial infection of 41.4%, with the least Aminu Kano Teaching Hospital (AKTH) has 29%. Among the Site of infection, 34.8% isolates were wounds Swabs (SSIs), urine (UTI) 31.2%, an least was nasal intubation 11.6%. Among pathogens isolated E. coli is the most prominent organism with 26.1% and the least Streptococcus pyogenes (7.9%). This study showed that the prevalence of Prolong hospital stay in Kano was 40.34%, AKTH 39.53% and MAWSH 28.91%. The prevalence of nosocomial infection was 34.3%. Gram-negative  bacteria were the predominant isolates

Comparative study of malaria parasite diagnosis: Microscopy and rapid diagnostic-testing

The conventional microscopy is the gold standard in laboratory diagnosis of malaria but time-consuming and requires a lot of training and expertise. However, Rapid Diagnostic Tests (RDTs) could be considered in endemic regions especially in poor settings, where there is shortage of power supply and less qualified personnel. This is a comparative study between Microscopy and RDTs. The study was carried out in two health facilities in Kano metropolis; Sir, Muhammad Sunusi Specialist hospital (SMSSH) and Bayero University Health Clinic (BUHC). A retrospective analysis was carried out in which the capacities of facilities for malaria parasites (MP) microscopy and RDT were assessed for the previous year. Later a prospective analysis was carried out by implementing a modified Lot Quality Assurance Sampling (LQAS) model over a period of 3 months. Giemsa stained Slides for Malaria Parasite Microscopy were sampled giving a total Slides of 432 and their corresponding blood specimens also sampled for the respective RDTs using SD Bioline Kits. The specificity of MP slide microscopy and RDTs in SMSSH and BUHC were 84% and 94% respectively while that of RDTs were 95% and 94% respectively. However, the sensitivity of MP microscopy in SMSSH and BUHC was found to be 95% and 94% and that of RDT was 72% and 78% respectively. Result showed that microscopic examination of malaria parasite is still the method of choice and also gold standard especially for confirmation of clinical diagnosis.Keywords: Malaria; microscopy; RDTs; sensitivity and specificit