Polymerase Chain Reaction for the Detection of Chlamydia psittaci in the Feces of Budgerigars

Abstract

The polymerase chain reaction (PCR) targeting the ompA gene of Chlamydia psittaci was evaluated for its ability to detect chlamydiae in fecal specimens of budgerigars as compared with isolation procedures using cell culture and embryonated egg inoculations. Several procedures for PCR template DNA preparation were compared so as to determine their detection levels for chlamydiae propagated in cell culture in the presence of fecal materials. Tween-20 and proteinase K treatments followed by centrifugation of the template DNA were found to be an appropriate procedure for DNA preparation for primary PCR. Subsequent nested PCR was shown to detect 4.8 IFU/ml or 84 particles/ml of chlamydiae. Chlamydiae in 50 fecal specimens from apparently healthy budgerigars were examined by nested PCR and several other methods. Nested PCR detected chlamydiae at a higher rate (12/50, 24%) than the isolation procedure in embryonated eggs (6/50, 12%). Primary PCR combined with the isolation procedure in cell culture gave a detection rate (5/50, 10%) similar to that of isolation from embryonated eggs. Detection rates by primary PCR (1/50, 2%) and in cell culture (0%) were inferior to the other procedures.

Chlamydophila psittaci DNA Detection in the Faeces of Cage Birds

“…psittaci had been transmitted from birds purchased in pet stores to humans. Takashima et al. (1996) examined Chlamydiae in 50 faecal specimens from apparently healthy budgerigars by nested PCR and detected Chlamydiae in 24% of the samples.…”

“…Polymerase chain reaction has been documented as a highly sensitive method for the detection of Cp. psittaci (Takashima et al., 1996; Trevejo et al., 1999). The technique offers advantages over other testing methods in that it is fairly rapid and does not require organisms to be present in large numbers or to be viable.…”

Isolation and Characterization of Chlamydophila psittaci Isolated from Laying Hens with Cystic Oviducts

“…Several PCR tests have been used for differential diagnosis of C. psittaci. Nested outer membrane protein polymerase chain reaction (OMP-PCR) tests are mainly performed for speciation of C. psittaci in poultry and occasionally for verification of real-time PCR results (16). In this study the nested OMP-PCR guaranteed specific diagnosis, in addition to simple handling and reduced contamination risk.…”

Development of a Lateral Flow Strip-Based Recombinase-Aided Amplification for Active Chlamydia psittaci Infection

“…detection, urethral swabs, choanal cleft swabs, pharyngeal swab, and cloacal swabs from animals or human are often used for DNA extraction and testing ( Holland et al, 1990 ; Dumke et al, 2015 ; Mina et al, 2019 ; Feodorova et al, 2022 ). One route of transmission of C. psittaci is a inhalation of aerosolized bird feces from the environment ( Szeredi et al, 2005 ), and fresh feces samples from animals are also used for detection of C. psittaci ( Takashima et al, 1996 ; Sachse et al, 2012 ; McGovern et al, 2021 ). Another advantage for the choice of fresh feces as a key specimen for detection of C. psittaci maybe that a feces collection is more convenient than a blood collection: rapid, harmless, and suitable for detection in filed.…”