CN113061666A - Detection method, application and system of cytomegalovirus after liver transplantation - Google Patents
Detection method, application and system of cytomegalovirus after liver transplantation Download PDFInfo
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Abstract
The invention discloses a detection method, application and a system of cytomegalovirus after liver transplantation, wherein the detection method comprises the steps of obtaining free DNA of a receptor sample after liver transplantation, and constructing and sequencing a free DNA sequencing library; comparing the sequencing data of the free DNA with a preset human reference genome, and setting the non-human source segment as a segment which is not successfully compared with the preset human reference genome in the sequencing data of the free DNA; comparing the non-human source segment with a preset cytomegalovirus genome, and setting a segment successfully compared with the preset cytomegalovirus genome in the non-human source segment as a cytomegalovirus segment; and calculating the number ratio of the cytomegalovirus fragments to the non-human fragments. The detection method provided by the invention has high sensitivity.
Description
Technical Field
The invention relates to the technical field of gene detection, in particular to a detection method, application and system of cytomegalovirus after liver transplantation.
Background
Liver transplantation is the only effective way to treat various end-stage liver diseases. The problem of infection after liver transplantation has been the main cause of long-term survival of patients, with incidence rates as high as 53% -80%. Cytomegalovirus (CMV) is a common opportunistic disease virus after liver transplantation, and cytomegalovirus infection is often one of the leading causes of increased incidence of complications and mortality. Patients who have received immunosuppressive therapy are at increased risk of developing cytomegalovirus infection, and patients who have undergone liver transplantation have a cytomegalovirus infection rate of 25% to 85% without prophylactic antiviral therapy, and 18% to 29% of patients develop cytomegalovirus disease. Therefore, the detection discovery of the cytomegalovirus is improved as early as possible, and the application of the cytomegalovirus in the diagnosis, prevention and treatment of patients can obviously improve the survival time of the patients and directly or indirectly reduce the incidence rate of other infections.
At present, the clinical detection of cytomegalovirus infection mainly comprises CMV antibody detection, pp65 antigen detection and CMV DNA detection, and specific detection methods comprise ELISA, qPCR technology and the like. However, CMV antibody test, pp65 antigen test, cannot simultaneously assess the severity of liver injury; the above examination method cannot exclude the condition that other organs (such as lung and kidney) of the human body are infected with cytomegalovirus; since the detection method has low sensitivity, erroneous determination is likely to occur.
Therefore, there is a need to provide a novel method for detecting cytomegalovirus after liver transplantation to solve the above-mentioned technical problems.
Disclosure of Invention
The invention mainly aims to provide a detection method, application and system of cytomegalovirus after liver transplantation, so as to solve the technical problem of low sensitivity of the existing detection method of the cytomegalovirus.
In order to achieve the above object, the present invention provides a method for detecting cytomegalovirus after liver transplantation, comprising the following steps:
obtaining free DNA of a receptor sample after liver transplantation, and constructing and sequencing a free DNA sequencing library;
comparing the sequencing data of the free DNA with a preset human reference genome, and setting the non-human source segment as a segment which is not successfully compared with the preset human reference genome in the sequencing data of the free DNA;
comparing the non-human source segment with a preset cytomegalovirus genome, and setting a segment successfully compared with the preset cytomegalovirus genome in the non-human source segment as a cytomegalovirus segment;
and calculating the number ratio of the cytomegalovirus fragments to the non-human fragments.
Preferably, the detection method further comprises:
obtaining original DNA of a receptor sample before liver transplantation, constructing and sequencing a receptor genome sequencing library, obtaining donor liver tissue DNA, and constructing and sequencing a donor genome sequencing library;
setting the fragments successfully aligned with the preset human reference genome in the free DNA sequencing data as human fragments;
and comparing the human source fragment, the sequencing data of the donor liver tissue DNA and the sequencing data of the original DNA of the receptor sample to obtain a donor human source fragment, and calculating the ratio of the donor human source fragment to the human source fragment.
Preferably, the step of comparing the human source fragment, the sequencing data of the donor liver tissue DNA and the sequencing data of the original DNA of the recipient sample to obtain a donor human source fragment, and calculating the ratio of the donor human source fragment to the human source fragment comprises:
comparing the original DNA sequencing data of the receptor sample and the donor liver tissue DNA sequencing data with the preset human reference genome respectively, and detecting to obtain a receptor DNA variation site and a donor liver tissue DNA variation site;
comparing homozygotes in the variant sites of the recipient sample with homozygotes in the variant sites of the donor liver tissue, and setting sites corresponding to different homozygotes as first analysis sites;
comparing the human fragment with the part of the donor liver tissue DNA sequencing data corresponding to the first analysis site, and setting the successfully-compared human fragment as the donor human fragment;
calculating the ratio of the donor human fragment to the human fragment.
Preferably, the first analysis site is a variation site with a mass value of greater than 30 and a sequencing depth of greater than 8X.
Preferably, the step of comparing the human source fragment, the sequencing data of the donor liver tissue DNA and the sequencing data of the original DNA of the recipient sample to obtain a donor human source fragment, and calculating the ratio of the donor human source fragment to the human source fragment comprises:
carrying out SNP chip detection on the original DNA of the receptor sample to obtain receptor SNP variation information, and carrying out SNP chip detection on the donor liver tissue DNA to obtain donor SNP variation information;
comparing homozygotes in the acceptor SNP variation information with homozygotes in the donor SNP variation information, and setting sites corresponding to different homozygotes as second analysis sites;
comparing the human fragment with the part of the donor liver tissue DNA sequencing data corresponding to the second analysis site to generate the donor human fragment;
calculating the ratio of the donor human fragment to the human fragment.
Preferably, the step of extracting free DNA from the recipient sample after liver transplantation, and performing free DNA sequencing library construction and sequencing comprises:
after liver transplantation, extracting free DNA of peripheral blood of a receptor sample, and constructing and sequencing a free DNA sequencing library.
The invention also provides an application of the detection method of the cytomegalovirus after liver transplantation in preparing a detection kit of the cytomegalovirus.
The invention also provides application of the detection method of the cytomegalovirus after liver transplantation in evaluation of liver cell damage.
The invention also provides a system, which comprises a processor, a memory, a communicator and a program of the method for detecting the cytomegalovirus after the liver transplantation, wherein the program of the method for detecting the cytomegalovirus after the liver transplantation is executed by the processor, and the steps of the method for detecting the cytomegalovirus after the liver transplantation are realized.
Compared with the qPCR detection and antibody detection mode in the prior art, the detection method provided by the invention has the advantages that the cytomegalovirus fragment in the receptor free DNA is detected, the sensitivity is higher, the cytomegalovirus can be detected in the body of a patient infected with the cytomegalovirus earlier, the clinical intervention can be carried out as soon as possible, and the survival rate of the patient can be further improved.
Drawings
FIG. 1 is a schematic flow chart of a method for detecting cytomegalovirus after liver transplantation according to an embodiment of the present invention;
FIG. 2 is a schematic flow chart of another embodiment of the method for detecting cytomegalovirus after liver transplantation according to the present invention;
FIG. 3 is a schematic flow chart of a method for detecting cytomegalovirus after liver transplantation according to yet another embodiment of the present invention;
FIG. 4 is a CMV infection amount curve graph using the detection method of cytomegalovirus after liver transplantation provided by the present invention and a conventional qPCR detection method;
FIG. 5 is a graph of a curve fit of the detection results of the detection method for cytomegalovirus after liver transplantation and the conventional qPCR detection method provided by the present invention;
FIG. 6 is a graph showing the content of donor humanized fragments obtained by the detection method of cytomegalovirus after liver transplantation according to the present invention;
FIG. 7 is a ROC curve of the detection method of cytomegalovirus after liver transplantation provided by the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In addition, the technical solutions in the embodiments of the present invention may be combined with each other, but it must be based on the realization of those skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination of technical solutions should not be considered to exist, and is not within the protection scope of the present invention.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by existing methods.
Referring to fig. 1, the present invention provides a method for detecting cytomegalovirus after liver transplantation, comprising the following steps:
step S1, obtaining free DNA of the receptor sample after liver transplantation, and constructing and sequencing a free DNA sequencing library;
free DNA (cfDNA) refers to extracellular DNA present in various body fluids and secretions such as blood, urine, saliva, etc., and is usually released upon apoptosis and necrosis of tissue cells. By detecting free DNA, genetic information of deep lesions or organs, fetuses, etc. can be obtained. Naturally, the free DNA in a broad sense also includes DNA of pathogens such as viruses and bacteria, and by detecting the free DNA, the infection of the pathogens in human body can be obtained.
The recipient is a patient receiving a liver transplant. The skilled person can select a suitable extraction method to extract free DNA from the recipient sample according to the actual situation. In this example, 10ml of peripheral blood of a patient after liver transplantation was collected using an EDTA blood collection tube, and plasma separation was performed within 2 hours to remove blood cells; 2ml of plasma was taken and free DNA in the plasma was extracted using a plasma free DNA extraction Kit (55114, QIAamp Circulating Nucleic Acid Kit, Qiagen, Germany). Since the free DNA itself is fragmented DNA, in this embodiment, the free DNA does not need to be broken, a conventional second generation sequencing library construction method is directly adopted to construct a sequencing library, and sequencing is performed by selecting a sequencing mode of a Hiseq X platform and PE150 to obtain at least 5G of sequencing data. Multiple reads are included in the sequencing data.
Step S2, comparing the sequencing data of the free DNA with a preset human reference genome, and setting the non-human source fragment as a fragment which is not successfully compared with the preset human reference genome in the sequencing data of the free DNA;
the reference genome is the genome sequence of the species, is the complete genome sequence which has been assembled, and is often used as a standard reference substance for the species. The predetermined human reference genome is a human genome predetermined by a person skilled in the art. Comparing the free DNA sequencing data with a preset human reference genome, wherein a fragment successfully compared with the preset human reference genome in the free DNA sequencing data proves that the fragment is from a human gene; the segment which is not successfully compared with the preset human reference genome in the free DNA sequencing data is proved to be a non-human segment, and specifically can be DNA of cytomegalovirus or other pathogens such as viruses and bacteria.
Step S3, comparing the non-human source segment with a preset cytomegalovirus genome, and setting the successfully compared segment of the non-human source segment with the preset cytomegalovirus genome as a cytomegalovirus segment;
the preset cytomegalovirus genome is a cytomegalovirus genome preset by one skilled in the art. And if the comparison is successful, the receptor patient is proved to be infected with the cytomegalovirus.
Step S4, calculating the quantity ratio of the cytomegalovirus fragments and the non-human fragments.
Specifically, the ratio of the reads of the cytomegalovirus fragment to the reads of the non-human fragment is calculated, and the ratio can reflect the load of the cytomegalovirus in the receptor body, namely the cytomegalovirus infection amount in the receptor body.
Compared with the qPCR detection and antibody detection mode in the prior art, the detection method provided by the invention has the advantages that the cytomegalovirus fragment in the receptor free DNA is detected, the sensitivity is higher, the cytomegalovirus can be detected in the body of a patient infected with the cytomegalovirus earlier, the clinical intervention can be carried out as soon as possible, and the survival rate of the patient can be further improved.
Further, the method for detecting cytomegalovirus after liver transplantation further comprises the following steps:
step S5, obtaining original DNA of a receptor sample before liver transplantation, constructing and sequencing a receptor genome sequencing library, obtaining donor liver tissue DNA, constructing and sequencing a donor genome sequencing library;
specifically, the recipient sample original DNA is the recipient's DNA prior to receiving a liver transplant. The method for extracting the original DNA of the recipient sample is not limited in the present invention, and in the present embodiment, 5ml of peripheral blood of the recipient is collected, and the DNA in the peripheral blood is extracted by a conventional phenol chloroform extraction method, and the DNA in the peripheral blood is the original DNA of the recipient sample. In this embodiment, 1ug of receptor sample original DNA is adopted to perform Covaris interruption, Ampure beads recovery and conventional second-generation sequencing library construction, a Hiseq X platform is selected, a PE150 mode is set (150 bp is respectively sequenced at two ends of the sequence), and sequencing is performed to obtain sequencing data of 10G to 20G.
The donor is a donor who provides a source of transplanted liver. In this example, when the donor liver was prepared before liver transplantation, a block of soybean-sized donor liver tissue was retained, homogenized by a grinder, and then extracted by a conventional phenol chloroform extraction method, wherein the donor liver tissue DNA was the DNA in the donor liver tissue. One skilled in the art can select suitable genome sequencing library construction and sequencing methods according to actual conditions, and in this embodiment, similar methods to those of the original DNA of the recipient sample are adopted.
And step S6, setting the successfully aligned fragment with the preset human reference genome in the free DNA sequencing data as a human fragment.
Step S7, obtaining donor human fragments through comparison according to the human fragments, the sequencing data of the donor liver tissue DNA and the sequencing data of the original DNA of the receptor sample, and calculating the proportion of the donor human fragments and the human fragments.
Referring to fig. 2, in an embodiment, step S7 specifically includes:
step S71, comparing the sequencing data of the original DNA of the receptor sample and the sequencing data of the donor liver tissue DNA with the preset human reference genome respectively, and detecting to obtain a receptor DNA variation site and a donor liver tissue DNA variation site;
by alignment with a pre-established human reference genome, sites of variation in the recipient and donor can be identified for use in distinguishing between individuals.
Step S72, comparing homozygotes in the variant sites of the receptor sample with homozygotes in the variant sites of the donor liver tissue, and setting sites corresponding to different homozygotes as first analysis sites;
homozygotes, also known as homozygotes, refer to genotypic individuals in which both alleles of homologous chromosomes are identical at the same locus, as opposed to heterozygotes. The first homozygote sites which are not identical are the alleles which are not identical in the recipient and donor. For example: the allele of the receptor is AA, and the allele in the donor corresponds to AA, i.e. AA is homozygote, AA is homozygote, and AA and AA are different homozygotes. A plurality of first analysis sites may be set in step S62. Preferably, the first analysis site is a variation site with a mass value of greater than 30 and a sequencing depth of greater than 8X. The sequencing depth is the ratio of the total base number (bp) to the Genome size (Genome) obtained by sequencing. Of course, the skilled person can set itself according to the actual need to select a suitable first assay site.
Step S73, comparing the human source fragment with the part of the donor liver tissue DNA sequencing data corresponding to the first analysis site, and setting the successfully compared human source fragment as the donor human source fragment;
specifically, the frequencies of the respective genotypes of the humanized fragments at the first analysis site determined in step S72 are analyzed by mutation detection software. Wherein the human fragment is considered to be derived from the donor liver tissue in accordance with the DNA sequencing data of the donor liver tissue.
Step S74, calculating the ratio of the donor human source fragment to the human source fragment;
the condition of liver tissue cell damage and death after transplantation can be reflected by calculating the proportion of the DNA from the donor liver tissue in the human source segment.
Referring to fig. 3, in another embodiment, step S7 specifically includes:
step S75, carrying out SNP chip detection on the original DNA of the receptor sample to obtain receptor SNP variation information, and carrying out SNP chip detection on the donor liver tissue DNA to obtain donor SNP variation information;
SNP (Single Nucleotide polymorphism) refers to a genetic marker formed by variation of a single Nucleotide in a genome, including transition, transversion, deletion and insertion. In this example, SNP chip detection was performed on 1ug of original DNA of the receptor sample to obtain the receptor SNP variation information.
Step S76, comparing homozygotes in the acceptor SNP variation information with homozygotes in the donor SNP variation information, and setting sites corresponding to the different homozygotes as second analysis sites;
the person skilled in the art can set the mass value of the second analysis site himself.
Step S77, comparing the human source fragment with the part of the donor liver tissue DNA sequencing data corresponding to the second analysis site, and setting the successfully compared human source fragment as the donor human source fragment;
step S78, calculating the ratio of the donor human source fragment to the human source fragment.
The condition of liver tissue cell damage and death after transplantation can be reflected by calculating the proportion of the DNA from the donor liver tissue in the human source segment.
The effective effect of the detection method of cytomegalovirus after liver transplantation provided by the invention is illustrated by the following specific examples:
cytomegalovirus detection was performed on 4 patients who received liver transplantation. The detection method provided by the invention and the conventional qPCR detection are adopted for 4 patients at the same time. Please refer to fig. 4, which is a CMV infection amount graph of the detection method of cytomegalovirus after liver transplantation and the conventional qPCR detection method provided by the present invention, wherein A, B, C and D are the CMV infection amount detection result and qPCR detection result obtained by the detection method provided by the present invention for 4 patients receiving liver transplantation, respectively, the abscissa is the number of days the patients receiving liver transplantation, and the ordinate is the ratio of the number of cytomegalovirus fragments to non-human fragments, and the qPCR quantitative value, respectively. The dotted line is the threshold for positive determination of CMV for this example. As can be seen from FIG. 4, before qPCR shows positive, the detection method provided by the present invention can detect the increase of the CMV DNA ratio by calculating the number ratio of the cytomegalovirus fragments to the non-human fragments, so as to prove that the detection method provided by the present invention has higher sensitivity and can detect CMV infection earlier than the conventional qPCR detection method.
Please refer to fig. 5, which is a graph of curve fitting between the detection method of cytomegalovirus after liver transplantation and the conventional qPCR detection method provided by the present invention. The figure shows the test results of all sampling points of the 4 patients, wherein the abscissa is the log10 transformation value of the qPCR test result, and the ordinate is the test value of the same sample by the method provided by the invention. If the patient has negative qPCR result, the point is not marked. As can be seen in FIG. 5, the fitting between the qPCR result and the result of the sequencing analysis method provided by the present invention is better, indicating that the method can better quantify the viral load in the patient.
Referring to fig. 6, a graph of donor human fragments using the method for detecting cytomegalovirus after liver transplantation provided by the present invention is shown, wherein the abscissa represents the number of days that a patient receives liver transplantation, and the ordinate represents the percentage of donor human fragments to human fragments. As the operation process inevitably causes certain liver injury, the proportion of the donor human source fragments in the early stage after liver transplantation is higher and gradually reduced along with the time. In the experiment, the proportion of the donor human source fragment of the patient is found to be increased when the proportion of the CMV DNA of the patient is increased in the later stage of liver transplantation, namely the cytomegalovirus fragment and the donor human source fragment are in positive correlation.
Please refer to fig. 7, which shows the ROC curve of the method for detecting cytomegalovirus after liver transplantation according to the present invention. The detection result of the conventional qPCR detection method is used for judging whether the CMV is positive, and ROC curve analysis is carried out on the CMV infection detection performance of the method, wherein the area under the curve (AUC) is 0.97.
The invention also provides an application of the detection method of the cytomegalovirus after liver transplantation in preparing a detection kit of the cytomegalovirus.
The reagent is prepared into the cytomegalovirus detection kit by adopting the cytomegalovirus detection method after liver transplantation. The application of the detection method of the liver transplanted cytomegalovirus in the preparation of the detection kit of the cytomegalovirus can refer to each embodiment of the detection method of the liver transplanted cytomegalovirus of the present invention, and details are not repeated herein.
The invention also provides application of the detection method of the cytomegalovirus after liver transplantation in evaluation of liver cell damage.
The application of the method for detecting cytomegalovirus after liver transplantation in assessing damage of liver cells can refer to various embodiments of the method for detecting cytomegalovirus after liver transplantation, and details are not repeated here.
Whether the tested patient is infected with the cytomegalovirus can be known by calculating the quantity ratio of the cytomegalovirus fragments to the non-human fragments, and the damage of the liver cells can be indirectly evaluated; by calculating the ratio of the donor human fragment to the human fragment, hepatocyte damage can be assessed directly.
The invention also provides a system, which comprises a processor, a memory, a communicator and a program of the method for detecting the cytomegalovirus after the liver transplantation, wherein the program of the method for detecting the cytomegalovirus after the liver transplantation is executed by the processor, and the steps of the method for detecting the cytomegalovirus after the liver transplantation are realized.
In an embodiment of the invention, the system may include a processor (e.g., CPU), a communication bus, a user interface, a network interface, and a memory. The communication bus is used for realizing connection communication among the components; the user interface may include a Display screen (Display), an input unit such as a Keyboard (Keyboard); the network interface optionally may include a standard wired interface, a wireless interface (e.g., WI-FI interface); the memory may be a high-speed RAM memory, or may be a non-volatile memory (e.g., a disk memory), and the memory may optionally be a storage device independent of the processor 1001.
Those skilled in the art will appreciate that the hardware configurations described above are not meant to be limiting of the system, and may include more or fewer components, or some components in combination, or a different arrangement of components.
The method for detecting the cytomegalovirus after the liver transplantation can refer to the embodiments of the method for detecting the cytomegalovirus after the liver transplantation, and the details are not repeated herein.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention, and all modifications and equivalents of the present invention, which are made by the present specification and directly/indirectly applied to other related technical fields within the spirit of the present invention are included in the scope of the present invention.
Claims (9)
1. A detection method of cytomegalovirus after liver transplantation is characterized by comprising the following steps:
obtaining free DNA of a receptor sample after liver transplantation, and constructing and sequencing a free DNA sequencing library;
comparing the sequencing data of the free DNA with a preset human reference genome, and setting the non-human source segment as a segment which is not successfully compared with the preset human reference genome in the sequencing data of the free DNA;
comparing the non-human source segment with a preset cytomegalovirus genome, and setting a segment successfully compared with the preset cytomegalovirus genome in the non-human source segment as a cytomegalovirus segment;
and calculating the number ratio of the cytomegalovirus fragments to the non-human fragments.
2. The method of detecting cytomegalovirus according to claim 1, further comprising:
obtaining original DNA of a receptor sample before liver transplantation, constructing and sequencing a receptor genome sequencing library, obtaining donor liver tissue DNA, and constructing and sequencing a donor genome sequencing library;
setting the fragments successfully aligned with the preset human reference genome in the free DNA sequencing data as human fragments;
and comparing the human source fragment, the sequencing data of the donor liver tissue DNA and the sequencing data of the original DNA of the receptor sample to obtain a donor human source fragment, and calculating the ratio of the donor human source fragment to the human source fragment.
3. The method of claim 2, wherein the step of comparing the human fragments, the sequencing data of the donor liver tissue DNA and the sequencing data of the recipient sample original DNA to obtain donor human fragments and calculating the ratio of the donor human fragments to the human fragments comprises:
comparing the original DNA sequencing data of the receptor sample and the donor liver tissue DNA sequencing data with the preset human reference genome respectively, and detecting to obtain a receptor DNA variation site and a donor liver tissue DNA variation site;
comparing homozygotes in the variant sites of the recipient sample with homozygotes in the variant sites of the donor liver tissue, and setting sites corresponding to different homozygotes as first analysis sites;
comparing the human fragment with the part of the donor liver tissue DNA sequencing data corresponding to the first analysis site, and setting the successfully-compared human fragment as the donor human fragment;
calculating the ratio of the donor human fragment to the human fragment.
4. The method of claim 3, wherein the first assay site is a variant site with a mass value of greater than 30 and a sequencing depth of greater than 8X.
5. The method of claim 2, wherein the step of comparing the human fragments, the sequencing data of the donor liver tissue DNA and the sequencing data of the recipient sample original DNA to obtain donor human fragments and calculating the ratio of the donor human fragments to the human fragments comprises:
carrying out SNP chip detection on the original DNA of the receptor sample to obtain receptor SNP variation information, and carrying out SNP chip detection on the donor liver tissue DNA to obtain donor SNP variation information;
comparing homozygotes in the acceptor SNP variation information with homozygotes in the donor SNP variation information, and setting sites corresponding to different homozygotes as second analysis sites;
comparing the human fragment with the part of the donor liver tissue DNA sequencing data corresponding to the second analysis site to generate the donor human fragment;
calculating the ratio of the donor human fragment to the human fragment.
6. The method of claim 1, wherein the step of extracting free DNA from the recipient sample after liver transplantation, constructing and sequencing a free DNA sequencing library comprises:
after liver transplantation, extracting free DNA of peripheral blood of a receptor sample, and constructing and sequencing a free DNA sequencing library.
7. Use of the method for detecting cytomegalovirus of any one of claims 1 to 6 after liver transplantation for preparing a cytomegalovirus detection kit.
8. Use of the method of detecting cytomegalovirus according to any one of claims 1 to 6 after liver transplantation for assessing liver cell damage.
9. A system comprising a processor, a memory, a communicator, and a program for detecting post-liver-transplantation cytomegalovirus stored in the memory and executable by the processor, wherein the program for detecting post-liver-transplantation cytomegalovirus when executed by the processor implements the steps of the method for detecting post-liver-transplantation cytomegalovirus of any one of claims 1 to 6.
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