Data will be analyzed using QuantaSoft software (Bio-Rad). The QuantaSoft program provides absolute quantities per microliter reaction mixture from given numbers of positive droplets and accepted droplets. The obtained values will be multiplied by 20 to calculate quantities in μl DNA extracts. If all the droplets have a positive signal, the template of DNA will be serially diluted to 10-2, and DD-PCR will be repeated as described.
The unites are expressed as copy numbers per microliter of DNA extract (T) will be calculated with the formula:
T=(-D)/Vp×ln〖(1-P/N)〗
Where ln is the base of the natural logarithm, Vp is the microliter of droplets, P is noted for the numbers of positive droplets, N is the total numbers of droplets, and D is the dilution rate of DNA templates. In this way the absolute number of bacteria and fungi can be estimated，