SARS-CoV computer virus was isolated from FRhK-4 cells cultured with a rectal swab specimen of a palm civet, but not from Vero E6 cells. syndrome, Rabbit polyclonal to APCDD1 coronavirus, palm civet, research The severe acute respiratory syndrome (SARS) epidemic emerged in 2003 in 6 municipalities in the Pearl River delta region in Guangdong, China. Early case-patients were more likely to be persons with occupational exposure to animals, such as animal sellers or restaurant cooks ( em 1 /em em , /em em 2 /em ). Tracing the source of infection has been complicated, given the sporadic nature of index cases without a clear history of contact with animals. After the World Health Business (WHO) declared the end of the SARS epidemic, 4 new cases of SARS were reported from December 16, 2003, to January 1, 2004, in Guangzhou in Guangdong Province. These cases were not linked to any laboratory accidents. All patients had a heat 38C, radiographic evidence of pneumonia, and serologic evidence of SARS contamination. Fever lasted from 6 to 18 days (median 7), no mechanical ventilation was required, and the clinical course of the disease ranged from 21 to 24 days with full recovery. All 4 patients had community-acquired infections without any apparent epidemiologic link. A total of 257 contacts, including 113 close contacts, of these patients were observed for 2 weeks, with no secondary transmission identified. These patients had mild symptoms and no secondary transmission, which was remarkably different from patients in the 2003 epidemic. Since potential reemergence of SARS leading to epidemic spread was possible, identification of the infectious source was a high priority. The S gene sequence of SARS-associated coronavirus (SARS-CoV) isolated from 2 of these 4 patients was Ki16425 found to be closely related to the sequence of computer virus isolated from palm civets ( em 3 /em ). However, 1 of these patients reported no contact with palm civets or other animals in the preceding 2 months. The second patient was a 20-year-old waitress from a restaurant that served palm civets as food ( em 4 /em em , /em em 5 /em ). Based on the virologic and epidemiologic findings, provincial officials took aggressive action on January 5, 2004, ordering a sweep through farms and food markets to eliminate any animals that might harbor SARS-CoV. No additional SARS cases have since been reported. This information highlights the necessity for investigating restaurants as a possible source of contamination, understanding that the computer virus can be transmitted from animals or environmental sources to humans, and clarifying the genetic basis Ki16425 of pathogenicity and infectivity of SARS-CoV from animal sources. Methods Specimen Collection Serial nasopharyngeal, fecal, and serum specimens Ki16425 of patients were collected at hospitals by Guangzhou Municipal Centers for Diseases Control and Prevention. When possible SARS was diagnosed in the waitress on January 2, 2004, serum, throat and rectal swabs were obtained from all 6 palm civets at the restaurant. It was reported that this animals were purchased from Xinyuan live animal wholesale market in Guangzhou. Serum samples from employees of the restaurant were obtained on January 4. Persons with positive results provided additional samples as needed. All specimens were stored at C80C. Laboratory Diagnosis and Direct Sequencing of Primary Specimens Serum samples were tested by enzyme-linked immunosorbent assay (ELISA), immunofluorescent antibody (IFA) test, and Western blot for specific immunoglobulin G (IgG) and IgM. Nasopharyngeal, throat, and rectal specimens were tested by reverse transcriptionCpolymerase chain reaction for polyprotein (P) and nucleocapsid (N) genes of SARS-CoV. Gene sequences were decided directly from initial samples. RNA was transcribed into cDNA (SuperScript, Invitrogen, Carlsbad, CA, USA) and subsequently used for PCR amplification. Complete spike (S) gene and whole genome sequencing of SARS-CoV computer virus was conducted by using 48 primer sets based on the sequence data of a Ki16425 SARS-CoV SZ3 isolate from palm civet ( em 6 /em ) and an ABI 3730 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). Assembled genome sequences were compared with those of the first computer virus isolates of human (TOR2) and animal (SZ3) origin. Any nucleotide (nt) differences.