Capacity building permitting comprehensive monitoring of a severe case of Lassa hemorrhagic fever in Sierra Leone with a positive outcome. Part 3
IgM and IgG endpoint titer determination: Sera were analyzed in 3-fold serial dilutions, starting at 1:50, in optimized LASV NP, GP1, GP2, Z protein combination ELISA, as outlined above. Reciprocal titers were calculated using mean +3 standard deviation (S.D.) cutoffs established with similarly diluted normal serum controls.
Piccolo®: The kinetics of fourteen serum analytes were analyzed daily using a Piccolo® blood chemistry analyzer (Abaxis, Inc., Union City, CA) with Comprehensive Metabolic Reagent Discs. Normal values were determined for a male in the age range of the patient using established clinical guidelines. Blood was collected in serum vacutainer tubes from patients and control donors and allowed to coagulate for 20 minutes at room temperature, followed by centrifugation in a tabletop centrifuge. The serum fraction was collected for analysis and aliquots were stored in cryovials at -20°C for future use.
Flow Cytometry: The kinetics of eleven serum cytokines were analyzed daily using an Accuri® C6 benchtop cytometer (Accuri Cytometers Inc., Ann Harbor, MI) with an eBioscience FlowCytomix Human Th1/Th2 11-plex Kit (Bender MedSystems GmbH, Vienna, Austria). Serum aliquots collected and frozen throughout the monitoring timeline were analyzed concurrently at the end of the study.
Presentation
Case 1: On September 1, 2010 the KGH LFW was contacted by the medical officer of Gondama Hospital in Bo district, Sierra Leone, concerning a suspected case of Lassa hemorrhagic fever. The patient was an eight-year-old male from Sembehun town, Malegohun chiefdom, Kenema district, Sierra Leone who had been first seen at the local health facility on August 30, 2010 (Figure 1). He presented to the health facility 2 days after the onset of symptoms with a history of persistent fever, headache, and profuse oral, nasal, and rectal bleeding. He was then referred to the Gondama Hospital where he could receive free medical care. After two days of treatment with anti-malarial and antibiotic drugs of unknown type and no improvement, the patient was referred to the KGH LFW as a suspected Lassa case. Upon arrival by ambulance at the KGH LFW on September 2, 2010, the patient reported having multiple symptoms including anorexia, malaise, headache, nausea, abdominal pain, loose black stools, hematemesis, dysuria, cough, sore throat and retrosternal pain. He presented with a body temperature of 38°C, a pulse rate of 140 beats/minute, and a respiration rate of 30. On examination, he was in obvious pain and lethargic. Bleeding from the mouth and nose was noted along with conjunctival injection, facial edema and hepatomegaly. During the first 24 hours after his admission to KGH LFW, he had multiple episodes of grossly bloody stools as well as hematemesis, hemoptysis and hemeturia. This patient was assigned the coded designation G-1180 upon initial diagnosis by NP LFI at KGH LFW, which will be used henceforth.