Enhanced Isolation of Mycoplasma pneumoniae from Throat Washings with a Newly Modified Culture Medium

Tully, Joseph G.; Rose, David L.; Whitcomb, R. F.; Wenzel, R. P.

doi:10.1093/infdis/139.4.478

Cited by 234 publications

(123 citation statements)

References 15 publications

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“…Nine species of AM and A laidlawii were isolated by IIF, in this study, but it has previously been demonstrated that all recognised species of AM can be detected in field isolates by the same method (BenCina, 1986). The use of IF in the identification of M. pneumoniae enhances significantly the demonstration of its isolation (Tully et al, 1979). Our findings regarding isolation of MG and MS and their demonstration are similar.…”

Section: Isolation and Identification Of The Field Mycoplasma Isolatessupporting

confidence: 77%

Mycoplasmaspecies in chicken flocks with different management systems

Benčina¹,

Mrzel

Tadina

et al. 1987

Avian Pathology

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SUMMARYChicken flocks hatched together but reared under different management systems were examined for mycoplasmas over a two-year period. On the farm A multiple-age flocks were reared in close contact. This farm had not been depopulated for over 15 years. On farms B, C and D single-age flocks were reared and the farms were depopulated every year. On farm A 218 birds from 20 flocks were tested: mycoplasmas were isolated from 202 (92.7%). On farms B, C and D 289 birds from seven flocks were tested: mycoplasmas were recovered from 55 (19.0%). On farm A the following mycoplasmas were identified: M. gallisepticum (46.9% of isolates), M. gallinarum (47.8%), M. pullorum (46.3%), M. gallinaceum (43.2%),M. iners (10.9%), M. iowae (8.3%),M. synoviae (51.7%), M. lipofaciens (23.8%) and M. glycophilum (16.7%). Furthermore, mycoplasma strains which do not belong to recognised species of avian mycoplasmas, Acholeplasma laidlawii and an unidentified Acholeplasma strain were also isolated. On farms B, C and D the isolates were identified as M. gallisepticum or M. synoviae. The only exception was one culture from which M. gallinarum was recovered. The differences between farm A and farms B, C and D regarding total mycoplasma isolation yields and incidence of their species are significant (/K0.01) and are attributed to the different management systems.

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Section: Isolation and Identification Of The Field Mycoplasma Isolatessupporting

confidence: 77%

Mycoplasmaspecies in chicken flocks with different management systems

Benčina¹,

Mrzel

Tadina

et al. 1987

Avian Pathology

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“…To prevent aspiration or swallowing of the flush, pressure was applied to the soft tissue between the ventral mandibles, forcing the tortoise's tongue into the choanae. The flush was collected in a sterile plastic container, and SP4 medium (Tully et al 1979) was immediately added to the nasal wash at a volume of 10% of the flush volume.…”

Section: Methodsmentioning

confidence: 99%

“…Culture and PCR techniques on nasal flush samples were similar to methods previously described (Brown et al 2001), except as follows. A 400-lL aliquot of the nasal flush sample was inoculated into 4 mL of SP4 broth (Tully et al 1979), 2 mL were filtered using a 0.45-lm filter to remove fungal contaminants, and 20 lL of each broth sample and the nasal lavage fluid were plated on SP4 agar. Broth cultures were analyzed for the presence of M. agassizii DNA based upon PCR amplification of a portion of the 16S rRNA gene .…”

Section: Methodsmentioning

confidence: 99%

Social behavior drives the dynamics of respiratory disease in threatened tortoises

Wendland

Wooding

White

et al. 2010

Ecology

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Since the early 1990s, morbidity and mortality in tortoise populations have been associated with a transmissible, mycoplasmal upper respiratory tract disease (URTD). Although the etiology, transmission, and diagnosis of URTD have been extensively studied, little is known about the dynamics of disease transmission in free-ranging tortoise populations. To understand the transmission dynamics of Mycoplasma agassizii, the primary etiological agent of URTD in wild tortoise populations, we studied 11 populations of free-ranging gopher tortoises (Gopherus polyphemus; n = 1667 individuals) over five years and determined their exposure to the pathogen by serology, by clinical signs, and by detection of the pathogen in nasal lavages. Adults tortoises (n = 759) were 11 times more likely to be seropositive than immature animals (n=242) (odds ratio = 10.6, 95% CI = 5.7-20, P < 0.0001). Nasal discharge was observed in only 1.4% (4/296) of immature tortoises as compared with 8.6% (120/1399) of adult tortoises. Nasal lavages from all juvenile tortoises (n=283) were negative by PCR for mycoplasmal pathogens associated with URTD. We tested for spatial segregation among tortoise burrows by size class and found no consistent evidence of clustering of either juveniles or adults. We suggest that the social behavior of tortoises plays a critical role in the spread of URTD in wild populations, with immature tortoises having minimal interactions with adult tortoises, thereby limiting their exposure to the pathogen. These findings may have broader implications for modeling horizontally transmitted diseases in other species with limited parental care and emphasize the importance of incorporating animal behavior parameters into disease transmission studies to better characterize the host-pathogen dynamics. Abstract. Since the early 1990s, morbidity and mortality in tortoise populations have been associated with a transmissible, mycoplasmal upper respiratory tract disease (URTD). Although the etiology, transmission, and diagnosis of URTD have been extensively studied, little is known about the dynamics of disease transmission in free-ranging tortoise populations. To understand the transmission dynamics of Mycoplasma agassizii, the primary etiological agent of URTD in wild tortoise populations, we studied 11 populations of free-ranging gopher tortoises (Gopherus polyphemus; n ¼ 1667 individuals) over five years and determined their exposure to the pathogen by serology, by clinical signs, and by detection of the pathogen in nasal lavages. Adults tortoises (n ¼ 759) were 11 times more likely to be seropositive than immature animals (n ¼ 242) (odds ratio ¼ 10.6, 95% CI ¼ 5.7-20, P , 0.0001). Nasal discharge was observed in only 1.4% (4/296) of immature tortoises as compared with 8.6% (120/1399) of adult tortoises. Nasal lavages from all juvenile tortoises (n ¼ 283) were negative by PCR for mycoplasmal pathogens associated with URTD. We tested for spatial segregation among tortoise burrows by size class and found no consistent evidenc...

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“…The culturing of M. pneumoniae was accomplished by a method adapted from that of Tully et al (20) and presented recently (C. J. Carlyn, A. L. Waring, C. K. Csiza, T. A. Halse, S. J. Wong, and R. J. Limberger, Abstr. 99th Gen. Meet.…”

Section: Methodsmentioning

confidence: 99%

Development of a Genomics-Based PCR Assay for Detection of Mycoplasma pneumoniae in a Large Outbreak in New York State

et al. 2001

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A genomics-based PCR method was developed and used to test specimens from patients involved in a large outbreak of Mycoplasma pneumoniae in a closed religious community in New York State. New P1 adhesin gene primers were designed to bind to 9 of 10 target sequences in the repetitive-element sequences obtained from the whole genome sequence of M. pneumoniae. This PCR method had a sensitivity of 0.006 CFU and a specificity of 100% for M. pneumoniae. The PCR was validated by testing a subset of patient samples by culture and comparing the results to those obtained by PCR. Of the initial 280 samples tested, 73 were positive by PCR and 22 were positive by culture. All samples positive by culture were also positive by PCR. Follow-up testing of selected patients 3 to 6 weeks after antibiotic treatment revealed that eight samples remained positive by PCR and that three samples remained positive by culture. Additionally, no nonspecific PCR inhibition was detected as a result of the specimen type, transport medium, or sample preparation methodology. The study demonstrates that the PCR described here is a rapid, sensitive, and specific method for the identification of M. pneumoniae and was helpful for the detection and monitoring of the outbreak.Mycoplasma pneumoniae is primarily a respiratory pathogen that is responsible for approximately 15 to 20% of all community-acquired pneumonias (8). The clinical presentation of patients with M. pneumoniae infection is not significantly different from that of patients with infections caused by other respiratory pathogens such as Chlamydia pneumoniae (22), so diagnosis of infection relies primarily on laboratory testing. M. pneumoniae is, however, a fastidious organism, requiring a laborious effort and 21 days or more for growth in culture. Likewise, serology is lacking in sensitivity, has questionable specificity (22), and is dependent on specific collection times relative to the onset of illness (5). DNA probes have also been used, but cross-reactivity between M. pneumoniae and Mycoplasma genitalium has been observed, and methods that use these probes also lack sensitivity (4).Numerous PCR approaches have been developed to provide a rapid, sensitive method for the detection of M. pneumoniae. PCR targets have included the P1 adhesin gene (3,4,5,12,17,21), the ATPase operon (11), genomic clones (2, 7), or a combination of these (1). The P1 adhesin gene is responsible for cytadherence, which is necessary for colonization and infection of host cells by M. pneumoniae (22). This gene is also an intriguing target for PCR because of its repetitive nature within the M. pneumoniae genome. Approximately 8% of this genome is composed of repetitive DNA elements with regions homologous to the P1 adhesin gene (10). Use of the entire genomic DNA sequence enabled us to design primers that would theoretically amplify the maximum number of target molecules, resulting in an improved sensitivity of the PCR for M. pneumoniae.A large outbreak of respiratory illness occurred in a closed religious communit...

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Enhanced Isolation of Mycoplasma pneumoniae from Throat Washings with a Newly Modified Culture Medium

Cited by 234 publications

References 15 publications

Mycoplasmaspecies in chicken flocks with different management systems

Mycoplasmaspecies in chicken flocks with different management systems

Social behavior drives the dynamics of respiratory disease in threatened tortoises

Development of a Genomics-Based PCR Assay for Detection of Mycoplasma pneumoniae in a Large Outbreak in New York State

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