Evaluation of Auramine O staining and conventional PCR for leprosy diagnosis: A comparative cross-sectional study from Ethiopia.

BACKGROUND: Diagnosis of leprosy mainly relies on clinical examination due to the inconsistent sensitivity and poor reproducibility of the current laboratory tests. Utilisation of alternative methods to the standard Ziehl Neelsen (ZN), Fite-Faraco (FF) and Haematoxylin and Eosin (H&E) staining procedures may eventually improve leprosy diagnosis. METHODOLOGY/PRINCIPAL FINDINGS: In this comparative study, the performance of the fluorescent Auramine O (AO) staining and polymerase chain reaction (PCR) was assessed with different skin samples using a combination of ZN, FF and H&E staining as the gold standard. AO, ZN, FF, H&E and PCR tests were performed on slit skin smears (SSS) and/or punch biopsies collected from 141 clinically confirmed leprosy cases and 28 non-leprosy skin samples. DNA was extracted from punch biopsies using two different methods with or without mechanical lysis. Sensitivities were 87.6%, 59.3% and 77% for H&E, ZN and FF, respectively, whereas it reached 65.5% and 77.9% for AO in SSS and tissue sections and 91.1% for PCR in tissue samples. Morover, samples with low bacillary index, sensitivity of AO staining (61.8%) was similar to FF (60%, p>0.05) and lower than PCR (86.6%, p<0.05). Sensitivity of PCR also increased (96.8%, p<0.05) when mechanical lysis was used during DNA extraction compared to enzymatic treatment alone (84.6%). CONCLUSIONS/SIGNIFICANCE: Our results showed that for diagnostic purposes, analysis of skin section is more sensitive than SSS, especially for samples with low bacillary load. AO staining on SSS and tissue sections was not significantly better than other routine diagnostic tests but considerably more user friendly. The sensitivity of PCR was higher than current standard methods and increased when combined with more efficient DNA extraction using mechanical and chemical lysis. Therefore, we recommend AO staining for the diagnosis of leprosy in lower health facilities such as health centres and district hospitals and PCR diagnosis at referral level and research centres.

An epidemiological study into the investigation of liver disease of unknown origin in a rural community of northern Ethiopia, 2005.

BACKGROUND: An outbreak of an unidentified cause of liver disease that claimed the lives of more than 45 people occurred in a rural community, in Northwestern Tigray. Despite the wider need for an urgent response there was no obvious explanation for the disease occurrence. OBJECTIVE: The aim of the study was to collect basic descriptive information and compare exposure status among case and control house-holds to better understand the illness and its pattern of occurrence so as to identify the sources, mode of transmission and possible cause of the illness. METHODS: The initial study principally involved combination of descriptive and analytic study designs. Combinations of quantitative and qualitative data were collected using semi-structured questionnaires. STUDY SUBJECTS: Samples of the affected and non-affected individuals of study village, key informants from various levels in the administrative system, and focus group discussions organized from both the affected and non-affected households of the study village were the subjects included for the study. RESULTS: The outbreak involved about 118 people, (78 males and 40 females) who were residents of the same village, in most, members of same family. More than half of the cases were under the age of 15 years. The outstanding clinical features were abdominal pain, rapidly filling ascites and hepatomegaly. The median duration of illness was six months (range; 1, 36 months). Of the 118 cases, 45 were deaths reported from the same community, during the period of illness with an estimated case fatality rate of 38%. Of the 45 deaths, 64.4% were children below 15 years of age. Qualitative information from both case and control communities showed difference in their water source. Eighty four percent of the dwellers in the affected village of Tsaeda Amba fetch water for their drink from an open shallow unprotected pond, Mai Habi-Tselam, Analysis of the data on exposure to the risk factor (case--control house-holds) revealed that users of the Habi-Tselam water source were almost 3 times more likely to develop the liver disease than those who used other water sources in the same village, OR = 3.14, 95% CI: 3.14 (3.12, 3.23). Age, nutritional status and long duration of exposure to an implicated source tended to influence the prognosis. CONCLUSION: The initial epidemiological investigations therefore gave an important insight as to the need to conduct further possible in-depth etiologic investigations, including clinical and toxicological studies, Study recommended an intermediary control measures including: banning the use of the implicated water source and provision of save and alternative potable water to the affected community.

An outbreak of veno-occlusive liver disease in northern Ethiopia, clinical findings.

BACKGROUND: Following the emergence of an unidentified fatal disease in Tahtay Koraro, rural farmers' village in Tigray, northern Ethiopia since 2001, a team of experts from Addis Ababa University Medical Faculty was assigned in November 2005, to investigate the problem in the field. The team consisted of an Infectious disease specialist, a Gastroenterologist, an Epidemiologist, a Pathologist, and a Microbiologist. The team was also joined by a group of Tigray Regional Health Bureauprofessionals. OBJECTIVES: To investigate the nature of unidentified liver disease, identify possible cause(s) and recommend specific interventions/treatment in order to save lives and stop further spread of the disease. METHODS: Data were collected from Clinical history, clinical evaluation of the affected cases in the area, relevant laboratory investigations, and histopathologic studies. RESULTS: The team evaluated sixty one of patients in the field (37 males and 24 females) with mean age of 27 years. The disease affected both gender and all age groups. Duration of the illness ranged from 1 month to 3 years with a median of 6 months. The illness was reported to have started with abdominal cramp in 75%, febrile syndrome in 77%, and abdominal distention in 63% of the cases. This was usually followed by jaundice, bleeding tendencies, and edematous states mainly in the form of rapidly filling ascites. Majority of the patients had similarly sick family members and/or lost family member(s) due to the same illness. Moreover, laboratory, sonography, histopathological, and toxicological studies revealed VOLD due to Pyrrolizidine alkaloid (PA) toxicity as described in the other sections of the supplement. CONCLUSION: From a detailed clinical and epidemiological investigation as well as histopathological and toxicological analyses, the disease was found to be fatal toxic hepatitis (veno-occlusive liver disease (VOLD). To our knowledge, this is the first report of VOLD from Ethiopia.

Veno-occlusive liver disease: a case report.

A case of veno-occlusive liver disease (VOLD) in a 12-years old Ethiopian boy is described The salient clinical features and gross and microscopic examination of biopsy material are reviewed. Veno-occlusive disease which occurs in the West Indies, East and West Africa, and India is an acute, subacute or chronic condition that affects the central and sublobular hepatic veins. In the West Indies (1) it is related to the consumption of bush tea made from plants that contain toxic pyrrolizidine alkaloids, such as Crotalaria and Senecio (2). Hepatotoxic compounds in Crotalaria, Senecio, Heliotropium and other composite plants can also enter the diet through the contamination of cereals with weed seeds. For example 28 of 67 patients died with veno-occlusive disease in central India after consuming a local cereal, gondli contaminated with the seeds of Crotalaria (3). Heliotropium Popovii has been implicated in outbreaks in villages in northwestern Afghanistan, with high mortality (4). The primary pathological change of hepatic veno-occlusive disease is sub-endothelial edema followed by intimal growth of connective tissue, with narrowing and occlusion of the central and sub-lobular hepatic veins. Atrophy or necrosis of liver cells, with consequent fibrosis leads to gross changes similar to those seen in cardiac cirrhosis, portal hypertension results. The present report, the first of it kind in Ethiopia describes a case of veno-occlusive liver disease in a 12-year old Ethiopian boy.