Table 1.

Samples Analyzed, Quantitative Polymerase Chain Reaction Results, Clinical Details, and Treatment Details

Time (months)relative to initialpresentation	Sample type	Panhemoplasma qPCR results:absolute no. of hemoplasma copies per PCR [GAPDH Ct value]	Clinical résumé and/or treatment
2	Serum	5.8 x 103 [30.1]	Pyrexia, pancytopenia, hepatosplenomegaly
5	Bone marrow	4.9 x 106 [21.8]	Pyrexia, hemolysis
5.25	Serum	1.2 x 105 [27.8]
6	Serum	Not detected [37.5]	DoxycyclineWithdrawal of prednisolone
7	Bone marrowa	4.7 x 104 [26.2] 6.1 x 105 [19.1]	Pyrexia, splenomegaly, hemolysis
8	Blood	<10 [21.7]	Doxycycline
8.5	Blood Bone marrow	<10 [22.7] <10 [21.6]	Doxycycline & moxifloxacin
9.5	Blood	<10 [22.1]
10.5	Blood	Not detected [21.5]
11.5	Blood Bone marrow	<10 [21.9] Not detected [19.7]
12.5	Blood	<10 [23.2]
13	Blood	Not detected [24.5]
14.5	Blood	Not detected [23.2]	Asymptomatic
14.75	Blood	Not detected [24.1]
15	Blood	Not detected [21.8]
16	Blood	Not detected [26.1]
18	Blood	Not detected [22.3]
19	Blood	Not detected [23.8]

Abbreviations: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was amplified as an internal control [1]. The threshold cycle (Ct) number was obtained by quantitative polymerase chain reaction (qPCR) for the GAPDH internal control. The qPCR results for samples of different origin cannot be directly compared with each other due to differences in dilution, collection medium, and volume used for DNA extraction. However, the hemoplasma copy numbers shown represent those present per PCR, which equates to number of copies per 5 μL of blood, bone marrow, or serum, assuming complete efficiency of DNA purification and PCR amplification. Stored samples were analyzed retrospectively. Because hemoplasma organisms are erythrotropic, PCR analysis of samples containing erythrocytes (blood and bone marrow) is usually preferred over that of serum samples. However, some serum samples generated positive hemoplasma PCR results in the current case report. We believe this likely reflects the very high numbers of organisms present in the blood at the time of serum preparation, meaning that detachment of even a small number of organisms from erythrocytes allowed their detection in the serum by PCR.

^aTwo different preservatives (Tryptic Soya Broth and Viral Transport Media) were used for bone marrow collection, and qPCR was performed on both, giving the 2 results shown.