背景:猫相关的嗜血支原体(血液原虫)被认为通过两种主要机制传播:(1)通过战斗直接传播和(2)由猫蚤(Ctenocephalidesfelis)传播。虽然C.felis的传输效率似乎很低,大多数手稿都集中在野生跳蚤中的血原虫的患病率,并报告了非常低(<3%)或高(>26%)的患病率。因此,我们旨在评估样品处理和PCR方法对C.felis血浆感染患病率的影响。
方法:对PubMed文章的系统审查确定了13份手稿(1,531只跳蚤/跳蚤池)符合纳入标准(对从猫收集的C.felis上的>1份血浆进行PCR)。使用ROBINS-E工具评估偏倚风险。在这些手稿的R中进行的荟萃分析发现,不洗涤样品和一组常见的16SrRNA引物首次发表在Jensen等人。2001年与血血浆患病率增加有关。为了评估洗涤对新收集的跳蚤的影响,我们评估了20个5个C.felis池的血浆状态,其中一半被洗了,一半没有洗。
结果:跳蚤冲洗并不影响血血浆的检测,而是扩增了螺血浆。用Jensen等人评估非特异性扩增。2001引物,对67例C.felis样品(34%先前报道的血血浆感染)进行PCR和测序。通过这种方法,仅在3%的样本中检测到血浆.在剩下的“血支原体感染”跳蚤中,PCR扩增螺旋体或其他细菌。
结论:因此,我们得出的结论是,在C.felis中的血浆感染是罕见的,未来的跳蚤流行研究应该对所有阳性扩增子进行测序以验证PCR特异性。有必要进一步研究猫相关的血血浆传递的替代方法以及C.felis维持血血浆感染的能力。
BACKGROUND: Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence.
METHODS: A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed.
RESULTS: Flea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining \"hemoplasma infected\" fleas, PCR amplified Spiroplasma or other bacteria.
CONCLUSIONS: Therefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary.