Plague Epidemic in the Kingdom of Naples, 1656–1658

Use of molecular probes and PCR for detection and typing of Leishmania—a mini-review.plifi cation of kinetoplast DNA as a tool in the detection and diagnosis of Leish-mania. rapid device for the serological diagnosis of Leishmania infantum infection in dog as compared with immuno-fl uorescence assay and Western blot. plague had spread to Sardinia and then to the cities and territories of Naples, Rome, and Genoa. Within the Kingdom of Naples, plague fi rst reached the town of Naples in the spring of 1656. Despite measures restricting population movement, by the summer of 1656, the disease had reached several provinces in southern Italy (1,2). Historical records indicate that the epidemic in Barletta, in southern Italy, developed after the arrival of a ship from Naples. On May 26, 1656, the ship Sant' Andrea arrived from Naples at the port of Barletta. However, after sanitary inspection, the ship was prevented from landing and obliged to depart, but this measure was not suffi cient to prevent the disease from entering the port. The Barletta epidemic peaked in October, after which the number of cases diminished; and on June 22, 1657, Barletta was declared free of plague. Of this city's original population of 20,000, the disease killed 7,000– 12,000 persons. It is hypothesized that throughout the Kingdom, the plague killed ≈1,250,000 persons (1,2). Since the 14th century, noble families of Barletta had been buried in tombs in underground tunnels of Sant' Andrea church. During restoration of the church in 2009, more underground tunnels containing many skeletons were discovered. It has been hypothesized that the church had also been used as a cemetery during the plague epidemic. During an inspection of the skeletons, 5 skulls of young persons were identifi ed and collected. For a negative control, the skull of a person buried in a tomb before the epidemic was also collected. The skulls were radiographed to identify unerupted teeth (Figure), which were then aseptically extracted. After classifi cation, each tooth was cut along a sagittal line to uncover the dental pulp, which was then hydrated in sterile phosphate-buffered saline (pH 7.2) for 48 h at 37°C. The DNA was extracted by using DNAeasy Blood and Tissue Kits (QIAGEN, Hilden, Germany) and by modifying the fi rst step, which was conducted overnight at 56°C with 600 μL of ATL buffer (QIAGEN) and 50 μL of proteinase K. To verify the presence of inhibiting substance, the control …