Jpn. J. Infect. Dis., 54, 12‑16, 2001

OriginalArticle

Evaluation of the AMPLICOR CMV COBAS AMPLICOR CMV Monitor andAntigenemia Assay f♭r Cytomegalovims Disease

TohruMasaoka* , Akira Hiraoka, Kensuke OhtaI , Noriyuki Tatsumi2, Shigeki Watanabe3,
Tomomitsu Hotta3, Hiromasa Yabe4, Shunichi Kato4, Atsushi Aikawa5, Takehiro Ohara5,
Akira Hasegawa5, Kazunari Tanabe6, Hiroshi Toma6, Akira Yasuoka7 and Shinichi Oka7

0saka Medical Center for Cancer and CaTdiovascular Diseases,
Nakamichi 1 ‑3‑3, Higashinari‑ku, Osaka 53 7‑8511,
1Department of Clinical Hematology and
2Department of Clinical and Laboratory Medicine, Osaka CiO, Universio7 Medical School,
Asahimachi 1‑4‑3. Abeno‑ku, Osaka 545‑8585,

3Department ofInlernal Medicine, Division ofHematology and Rheumatology and
4Department of Pediatrics, School ofMedicine, Tokai Universioi
Shimokatsuya 143, Isehara, Kanagawa 259‑1193,
5Tob,o Women女Medical College Kidney Center,

Kawadacho 8‑1, Shinjuku‑ku, Too/0 I 62‑8666,6Department ofNephy10logy. School ofMedicine, Toho Universio!,

Omori‑nishi 6‑ll‑1, Ohta‑ku, Too/0 143‑8541, and
7AIDS Clinical Center, International Medical Center ofJapan.
Toyama I‑21‑I, Shinjuku‑ku, Tokyo 162‑8655, Japan(Received September 12, 2000. Accepted January 3 1 , 2001)
SUMMARY: The AMPLICOR CMV (qualitative DNA assay by PCR), CO凸AS AMPLICOR CMV Monitor(quantitative DNA assay by PCR),andantigenemia. assay were tested for their ability to diagnose cytomegalovirTs(CMV) infection in 1 1 5 immunocompromised patlentS･ The AMPLICOR qualitative assay and the antigepeヮlaassay were positive for all nine patients with a clinical diagnosis of CMV disease. The AMPLICOR quantltatlVeassay was negative for one of the nine patients. In 1 06 patients without CMV disease, the AMPLICOR qualitative
test was positive in 22, the quantitative test was positive in 23, and the antigenemia test was positive in 55

patients. The AMPLICOR qualitative and quantitative assays had specificities of 79% and 78% in patients

without CMV disease, while that of the antigenemia assay was 48%. Diagnostic eLrlCiencies were 79% for the
AMPLICOR qualitative assay, 69% for the AMPLICOR quantitative assay, and 48% for the antigenemia assay.
All three tests yielded positive results before, or at the same time as, the onset of CMV disease in most cases,
which suggests they can be used to predict disease before the onset of symptoms. During antiviral treatment, test

results tended to decrease quantitatively and finally became negative; negative results were followed by remissionof symptoms. This suggests that the AMPLICOR quantitative assay and the antigenemia assay could be usefulfor monitoring therapeutic efrlCaCy. The AMPLICOR qualitative and quantitative assays, as well as the antigenemia
assay were considered effective for all of the following: diagnoslng CMV disease, predicting the onset of disease,and evaluatlng the effectiveness ofantiviral chemotherapy. The antlgenemia assay was at times dilrlCult to performin the case of severely neutropenic patients, whereas the AMPLICOR assays could be used in such cases.

Calif., USA) (7). It has been reported that the sensitivity of

INTRODUCTION

the AMPLICOR qualitative assay could be adjusted byestablishment of a suitable cut‑off point (8). In order toCytomegaloviruS (CMV)infection may cause seriousdisease
in immunocompromised patients. Antigenemia testlng anddetermine the sensitivlty and specificity of these tests and tocompare their performance to that of the antlgenemia assay,polymerase chain reaction (PCR) are now considered promi5‑
ing methods of diagnosing of CMV (ト3). The antigenemla

We conducted a clinical study in patients who underwent bone

assay has been recomended with several guidelines f♭r bonemarrow transplantation (BMT) or kidney transplantation (KT)marrow transplant patients (4,5)･ However, S9Ple reports haveand in patients with acquired immunodeficiency syndrome

suggested that both me仇ods may be too sensltlVe f♭r diagnos‑(AIDS).

1ng CMV infection, predicting disease onset, and monitonng
the clinical course of disease (6).

MATEIRALS AND METHODSThe AMPLICOR qualitative and quantitative assays weredeveloped by Roche Molecular Systems, Inc. (Pleasanton,

Patients and samples: A total of 1 1 5 patients were reglSteredas follows: BMT: 54 patients hospitalized for BMT. KT: 39*Corresponding author: Tel/Fax: +81‑6‑69811153 I, E‑mail:patients hospitalized fわr KT. AIDS: 22 patients with CD4

massanto@bh. mbn. or.jp

1ymphocyte counts of less than 100/LEI. Infomed consent

12was obtained from all patients. Blood samples were collected

detection were automatically perfわrmed with a CODAS

from patients with BMT and KT at weekly intervals for 12‑

AMPLICOR Analyzer manufactured by Roche Molecular

1 6 weeks, startlng the week befわre transplantation. Blood

Systems, Inc.

samplesfrom AIDS patients were collected at weekly intervals
for 12‑ I 6 weeks. Eight patients with symptoms such as fever

The COBAS AMPLICOR Analyzer calculates the CMVDNA levels in the test specimens by comparlng the CMVor cough were followedfurther at a physician's request.

signal to the qS signal fわr each specimen. Results werelaboratory. Serum was separated and tested by PCR on theexpressed as number of copies/ml. The CODAS AMPLICOR
Analyzer reports the result as invalid for any sample with asane血y the blood was collected. Similarly, white blood cells

qS optical density outside or an acceptable range, which is

Blood samples were kept at 2‑80C during transport to the(WBC) W?re isolated from whole blood and tested in theautomatically programmed into the Analyzer. The limit ofantlgenemla assay On the day or collection.

detection or this assay is 400 copies/ml.

Antigenemia test: This assay was performed with a CMV
Kogen "Mitsubishi" kit, manufactured by Yuka Medias Co.,AMPLICOR CMV: This is a qualitative PCR test fわr thedetection of CMV DNA in serum and was performed accord‑
was mixed with 500 LEI of extraction reagent and incubated

Ltd･ (Ami, Ibaraki)･ AssTys were conducted according to the
manufacturer's instructlOnS (9). A 4‑ml sample of EDTA ‑at lOOoC for 30 min. The mixture (50JLl) was transferred

anticoagulated venous blood was mixed with 1 ml dextran

lng tO the manufacturer's instructions. Briefly, 50 LL I serum

into the PCR tube containlng all components necessary forsolution. A鮎r sedimentation f♭r 15 min at 30oC, the super‑

amplification, plus an intemal control (IC). The IC was anatant was collected and centrifuged at 150 G for 6 min. The

non‑infectious plasmid with primer binding sequences iden‑

red blood cells were lysed with 3 ml of erythrolysIS solution

tical to and signal sequences distinct from the target DNA.

f♭r 5 min on ice.

The IC was added to permit the identi丘cation of processed

The pellet was washed, centrifuged, and resuspended at a

specimens containlng Substances that may have interferedcell concentration of 1.5 × 1 06/ml. One hundred microliters

with PCR amplification. Amplification was performed on a
GeneAmp PCR system 9600 thermal cycler manufacturedof suspension was spotted onto a slide with a cytocentrifuge,

by Applied Biosystems (Foster, Calif., USA). After ampli丘ca‑

stained by an immunonuorescence assay with a monoclonaltion, nucleotide sequences were hybridized to oligonucleotide‑

antibody agalnSt CMV pp65 antigen. The slides for eachand the suspension was then driedand fixed. The slides were

coated microwell plates. The probe‑bound ampli丘ed products

specimen were prepared in duplicate. The presence of positive

were detected colorimetrically with an avidin‑horseradish

cells/1.5 × 105 WBC was considered as a positive result. The

peroxidase conjugate and 3,3 '5,5 '‑tetramethylbenzidine

results are calculated by taking the average of two slides.

Criteria for climical diagnosis of CMV disease: The(TMB). Absorbance was measured at 450 nm on a NovaPath
MICROPLATE READER manufactured by Bio‑Rad Labo‑

criteria fb∫ clinical diagnosis of CMV pneumonia, enteritis,

ratories (Hercules, Calif., USA). Samples with an absorbanceretinitis, systemic infection, hepatitis,and other CMV diseases

value or ≧0.35 were considered positive. Samples with an

were determined by a committee that relied pnmarily on

absorbance value of<0.35and IC withanabsorbance of≧0.35

criteria previously reported by Ljungman and Plotkin (I 0),

were considered negative. When the absorbance for bothas shown in Table I. Patients were classifiedwith systemic

CMVand the IC were below 0.35, the results were considered

infection when they presented with symptoms suspIC10uS Of

invalid. ln such cases, it is generally recommended that

CMV infectionand when they tested positive for CMVantigen
or CMV DNA by in‑house testlng.

another aliquot of the original specimen be processed, or that

Period of study: This study began in September 1998 anda new specimen be collected. The limit or detection of thiswas completed in August 1999.

assay was 1000 copies/ml.

COBAS AMPLICOR CMV monitor: This quantitative
PCR test fb∫ CMV DNA in senlm Was perf♭med according

RESUIJTS

to the manufacturer 's instructions. Two hundred microlitersof serum was mixed with 600 LEI oflysIS reagent containing a

Patients: Ofthe 1 1 5 patients, 9 were diagnosed with CMV

known number of quantitation standard (QS) DNA molecules.

disease (Table 2). Four patients were diagnosed withThe DNA was preclpltated with isopropanol, washed with
70% ethanol, and suspended in the Monitor Specimen Diluent.

pneumonia, 2 with retinitis, 2 with gastroduodenal ulcer, 1
with hepatitis, and 1 with systemic disease; 1 patient

Fifty microliters of processed samples were added to 50 LLI

presented with pneumonia and retinitis. The remainlng 1 06of the master mix for amplification. Amplification andpatients were free of CMV disease throughout the study.

Table 1. Criteria for diagnosis of CMV disease
CMVDisease

ﾆ匁

CMVPtleumOniat

ﾄf匁F匁w2

6

W7E%

CMVEnteritis

W6Vf
ﾖ友匁x.&&F
ﾖ匁④HistologyorCMVbybiopsy

CMVRetinitis

G

CMVSystemicfWfW(tInfection

ﾆ

⑦

vV觚&

ﾆﾖ

ﾆf

DFWFV7F柳踟$4ﾕf

CMVHepatitis

&踟&ﾖ

ﾆ

ﾆv

觚Vﾖ

ﾇ

冶t

ﾖVﾆV

7儲

F

t$6ﾆ匁

ﾗ8t5

ﾇ7儲

6友庸T4ﾕf'

F

ﾗ77V6

7

ﾆﾆ

W

F

&&薮

7

Vﾆ6W"

ﾆf匁F匁rF殆VXt$ﾆWVｶ

7友

蹤没V踟$4ﾕdD

庸W&gV

譁

F柳竊VﾆWf

V譁

t5F

匁&ﾆ

F柳踐d

ﾖ&

7友

V譁

B

5C"ﾄ

ﾅC2

F

ﾆ

v

ﾆFWFV7F柳踐d4ﾕb③exclusionorotherviralhepatitis

OthersVW

ﾆ友

I BAL: Broncho‑alveolar lavage

t

觚

友

t%

&V

F友

t4

&v

儲

F

ﾖ

襷FWFV7F柳踐d4ﾕf也F

2AST:Aspartate aminotransferase ･1ALT:Alanine aminotransferase

13

F

&v

Table 2. Patients with CMV disease

CaseNumber
1

F坊蹠2

忍2

2

忍2

3

CMVRetinitis

忍2

4

Esophag￠alulcer

忍2

5 噺芳

CMVPneumonia

膝&6 噺芳觚膝&7 噺芳觚膝&
8

覽ﾖ

9

ﾆ

蹤

F柳

CMVPneumonia

ﾆ

蹤

F柳

CMVEnteritis

ﾆ

/4

覽ﾖ

CMVDisease

CMVPneumonia

蹤

uG&

/4

F柳

ﾆ

uG&

CMVSystemiclnfection蹤F柳

CMVRetinitis,CMVPneumonia

ﾆ

蹤

F柳

CMVHepatitis

Sensitivity for CMV disease: The sensitivity, specificity,

without CMV disease, the AMPLICOR qualitative test wasand diagnostic e銃ciency of the respectlVe tests are Surrmarized

positive in 22, the AMPLICOR quantitative test was positivein Table 3. Both the AMPLICOR qualitative assay and thein 23, and the antlgenemia assay was positive in 55 patients.

antigenemia assay yielded positive results for at least one blood

The respective specificities for CMV disease of the
AMPLICOR qualitative assay, AMPLICOR quantitative

sample fわr each of 9 patients with CMV disease. However,the AMPLICOR quantitative assay results were negative in 1

human immunodeficiency virus (HIV)‑positive, had an esoph‑ageal ulcer, and was diagnosed with CMV retinitis after theassay, and antlgenemia assay were 79%, 78%, and 48%.
Diagnostic efrICiency: The diagnostic efficiency of eachtest was calculated by the following formula: sensitivity (%)/
I 00 X specificity (%)/1 00. Respective efficiency percentages

end of the observation period. No treatment was administered

were 79%, 69%, and 48% for the AMPuCOR qualitative,

during the observation period. The AMPLICOR qualitative
assay was positive only once during the obseⅣation period.

AMPLICOR quantitative assay, and antlgenemia assay.
Time course from CMV detection unta disease diagnosis:

The antlgenemia assay was positive from the second week,

The time course ofCMV disease and assay resultsare shoⅥm

but the number of positive cells was consistently less thanin Table 4. In 9 patients, 12 episodes ofCMV disease wereobserved. The AMPLICOR qualitative test was positive inpatient (case 3) among these 9 patients. This patient was

10/1.5 × 105 WBC during weeks 2‑12.

The sensitivities of the AMPLICOR qualitative,

10 episodes. ln 6 0rthe 10 episodes, this test was positive at

AMPLICOR quantitative, and antigenemia assays for CMV
disease was 100%, 89%, and lO0%, respectively.

least 1 week before the onset of disease. The AMPLICOR

Specificity for CMV disease: Among the 106 patients

episodes, it was positive before the onset of disease. The

quantitative test was positive in 9 episodes. In 6 or the 9

Table 3. Sensitivity, Specificity and diagnostics efrlCiency of three assays

Assay

ﾕfF V 6R

6W2

6友庸T v F庸2 Sensitivity(%) 疲

ﾕfF V 6R ﾄ f6 672

1)AMPLICORqualitativcassay 涛2)AMPLICORquantitativeassay 塔3)Antigenemiaassay涛

6友庸4 v F庸R SpecirlCity (%) 認 v踟7F

100
89

loo

#ィ
3

鉄SS

79
78

都
田

48

鼎

'

踪ﾈﾇF庸R

Vﾖ6坊 窒R

Diagnostic efficiency was obtained as sensitivity(%)/1 00 × specificity(%)/I 00

Table 4. Time course of CMV disease (Weeks)
Case

W&柳F

$4ﾕb

F

V

6R

PeriodofAntiviral Therapy
+

1

sB

1‑6

白ﾓ2ﾃ

釘ﾓ
13‑20
ト27
冢one

2

25‑27
3

3‑7

迭ﾓb

6

澱ﾓr

7

B

8

rﾓCb

9

湯ﾖﾆﾂ

7‑9

ll‑13

覲

ﾓC

迭ﾃ

0‑4,9‑132

24‑29,41‑47

ﾄ

匁

覲

匁

覲

匁

0,3,5‑6,8

B

14

0‑20

ﾓ#

s2ﾃR

5‑7

6s

"

rﾃ#蔦3

ﾓ#"

0

0,7

迭ﾓb

none

7‑9

釘ﾓRﾃrﾃ0‑1224‑25,27,29, 41‑43,45‑47

ﾃ

7,9‑10Rﾃ
津#8,9
0‑4,9‑13

蹤没V觀ﾖ

辻

23‑27

0,7

覲

V

+

s"ﾃBﾃ

s"ﾃBﾓR

2ﾃ

迭ﾓbﾃ

ﾕ

ﾓ#2
0‑1,4‑9ll,13,16,1.9
ﾓbﾃ

迭ﾃrﾓ

ﾃ3Rﾃ3

0‑4,6‑9,ll‑14

F庸R

辻

湯ﾓ

0‑12,14

ﾓ3

ﾆ友

0,4‑6

5‑7

8

迭ﾃrﾓ

13‑14

25‑47

s2ﾃ

0,1,7

唐ﾓ

V

+

0‑2,4‑20

匁

'

24‑29

ﾃBﾓR

5‑6

8‑9

ﾈuD4

ﾃ"ﾓB
ﾓ#21,6‑9
none

none

ﾓR

5

0,4‑6

4‑27.

ﾓ#

4

ﾕ

辻

6s

ﾃ3"ﾂ

0‑4,7‑9,13‑142

0‑3,8‑12
2ﾃ

3Rﾓ3

B

0‑10

24‑26,28,31, 34,40‑47

ﾃBﾓRﾃ蔦

ﾆﾆﾂ

3,6‑8,ll‑14

Table 5. Test resultsfor patients without CMV disease
Numberofcases Tbta1106

Tcstrcsults

AMPLⅠCORqualitatiVe

ﾕ

T4

'

V

蹤友

庸R

50

辻

‑

辻

20

調

+

調

32
I

辻

‑

調

Antigenemia

調

+

辻

I

調

‑

調

2

辻

+

調

Table 6. Frcquencics of positive on antlgenCmia assay

32cascsnon‑CMVdiscascgroupcasesofCMVdiseasegroup

Frequencyorpositivetest 皮Vﾖ&W&&6

duringobscrvationpcriod(times)囘uringobseⅣationperiod(times)

1
2

6W2３

bゴ
免ﾂ

3

ッ

Frcquencyofpositivetest 皮Vﾖ&W&

R

1

B紕R

R

R

3

R

R

4

2

R

5

R

5

"

R

6

R

6

R

≧7

6W2３

R

2

4

≧7

f6

R
釘イB紕R

than five positive cells/1.5 X 105 WBC as shownin Table 6.

antlgenemia assay was positive in all 12 episodes. In 8 of the

1 2 episodes, it was positive before the onset of disease. These0ther aspects: Due to leukocytopenia, the antlgenemia

findings suggest that these tests may be effective for preemp‑

was not tested during the 1‑2 weeks after BMT in 3l of54

tive or early diagnosis of CMV disease.
CompletioA Of antiviral therapy and negative test
results: h cases 1, 4, 5, 6,and 9, all three tests became negative

patients. The AMPLICOR qualitative and quantitative assays
were tested during that period. The AMPLICOR quantitative
assay yielded invalid QS results several times in 6 patients. It

a鮎r several weeks of antiviral ther叩y and aRer the clinicalappears thataninterfering substance was present in the blood

symptoms had also subsided. In case 8, all three tests remained

samples of these patients. Although heparin is known tointerfere withPCR, we did not use heparinfor blood collection.

positive aRer acyclovir and ganciclovir administration andthey became negative a洗er fbscamet administration, which

induced a remission of symptoms. In case 7, the onset ofCMVDISCUSSION

disease occu汀ed just prlOr tO the end of the observation

Forthe diagnosis of CMV disease, the AMPLICOR qualita‑

period of 14 weeks. Thus, tests were not perf♭med on thisQuantitative difference between true and false positive

tive assay had a sensitivity of 100%, a speciflCityof79%, anda diagnostic efnciency of 79%. The AMPLICOR quantitative
assay had a sensitivity of 89%, a specificityof78%, and adiagnostic efficiency of 69%. The antlgenemia assay had asensitivity of lO0%, a specificityof48%, and a diagnostic

tests: Results of all three tests for patients without disease
are shown in Table 5. The false positive rates were 20.8% fortive assays thus appear to yield higher specificityand higher

the AMPLICOR qualitative assay, 2 I.7% for the AMPuCOR

diagnostic efficiency.

quantitative assay, and 5 1.9% for the antlgenemia test. For
the AMPLICOR quantitative assay, the average copies/ml was
3,746 from 3 1 samples in the group with CMV disease and1,948from 50 samples in the group without CMV disease.
The number of copies between the two groups was statisticallydifferent (P ‑ 0.029).

Boivin?t al･ (1 1) reported that in HIV infected patients, theantigenemla assay Was Very Sensitive (96%), yet less specificpatient during the course of the study. In case 2, tests became
negative after the first treatment; at the time of the second

treatment, the AMPuCOR qualitative and quantitative assays
were already negative.

efficiency of 48%. The AMPLICOR qualitative and quantita‑(83%) than the AMPLICOR qualitatiye assay (sensitivity:

65%, specificity: 92%) Or a PCR testuslng leukocyteswith acut‑off point of 690 copies /105 cells (sensitivlty: 65%,
specificity: 94%). Tong et al. (12) tested eight types of
assay, 1mCluding an antlgenemia assay and the AMPLICOR

As regard to the antlgenemia assay, the average number of

positive cells/I.5 X 105 WBC was 13.I for 65 samples in thequalitative test, in 37 kidney transplant patients. They reported

group with CMV disease and 4.5 for 1 50 samples in the group
without CMV disease. The number of cells between the twothat the AMPLICOR PCR produced the best results andfurthermore that it predicted CMV disease 2‑6 days before

groups was statistically different (P ‑ 0.001 ).
Among the 32 patients who received false positive results

the onset of symptoms.

by the antlgenemia assay only, positive results occu汀ed 1‑2

antlgenemla assay Was elevated by setting the cut‑off point at

Pellegri.n et all (13) reported that the specificity of thetimes in 84% of the patients, and the number of positive cells

five positive cells. In the present study, we observed similarly

was lower than 4/1.5 X 105 WBC cells in 92% of the positive
samples. In the patients with CMV disease, positive resultshigh sensitivities in all three tests, as reported. When the
AMPLICOR qualitative assay wasperfomed with the cut‑were observed more frequently than four times with more

offPolnt Set at 0.35 absorbance, we observed similarly high15

specificity. In the antlgenemia assay, the result was judged

5. Guideline Committee of Japanese Society for Hemato‑

positive with a positive cell /1.5 × 105 WBC. Ifa suitable

poietic Cell Transplantation (1997): Guidelines f♭r

cut‑off point could be established, a higher grade of specificity
might be obtained. For example, when a cut‑off point of 5/

prevention of cytomegaloviruS infection in bone marrowtransplant recipients. Japanese Society for Hematopoeitic
Cell Transplantation (in Japanese).

1.5 × 105 was set in the case of the antlgenemia assay in thisstudy, the specificityincreased from 48. 1% to 77.4%.

6. Patel, 良., Smith, T. F., Espy, M., Wiesner, 良. H., Krom,

All three tests produced positive results in 7 of 9 patients

R. A･, Portela, D. and Paya, C. Ⅴ (1994): Detection ofand 8 of 12 episodes. These results were obtained before, or at

cytomegalovirus DNA in sera of liver transplant recipients.

the same time as, the onset ofCMV disease. The AMPLICOR

CMV disease. This suggests that early diagnosis may be

J. Clin. Microbio1., 32, 1431‑1434.
7. Hiyoshi, M., Tagawa, S., Takubo, T., Tanaka, K., Nakao,T., Higeno, Y., Tamura, K., Shimaoka, M., Fujii, A.,
Higashihata, M., Yasui, Y., Kin, T., Hiraoka, A. and
Tatsumi, N, (1997): Evaluation of the AMPLICOR CMV

possible. Aitken et al. (14) reported that 14 out of52 kidney

test f♭r direct detection or cytomegalovims in plasma

qualitative test, the AMPLICOR quantitative test, and theantlgenemia assay all produced positive results at 1.9 weeks,
1.8 weeks, and 2.4 weeks, respectively, befわre the onset oftransplant patients with CMV disease tested positive by the

specimens. ∫. Clin. Microbio1., 35, 2692‑2694.

AMPLICOR assays 12.5 days befわre the onset of CMV

8. Long, M., Drew,し., Miner, 良., Jekic‑McMullen, D.,disease･ In BMT patients, Boeckh et al. ( 1 5) reported that the

Impra.im, C･ and Kao S･‑Y･ (1998): Detection ofcytome‑

antlgenemia assay guided preemptlVe treatment With goodtiming･ Einsele et al･ (1 6) also reported that PCR monitor享ng

galovlruS in plasma and cerebrosplnalfluid specimensfrom human immunodeficiency virus‑infected patients

reduces the severlty Of CMV disease as well as the duratlOnby the AMPLICOR CMV test. ∫. Clin. Microbio1., 36,of antiviral therapy･ The Japanese Society for Hematopoietic

2434‑2438.

Cell TransplantatlOn (5) recomended the antigenemia assay

9. Kurihara, T., Hayashi, J., Matsuoka, T. and lto, A. (1995):
HCMV pp65 antlgenemia assay uslng in direct alkaline

in the 1997 Japanese guidelines f♭r BMT. In the Centers f♭r

Disease Control and Prevention (CDC) guidelines fわr BMT

phosphatase stainlng method. Biomed. Res., 16, 125‑129.

in 2000, Dykewitz et al･ (4) recom甲ended monitoring patients

10. Ljungman, P. and Plotkin, S. (1995): Workshop on CMV

weekly with either the antlgenemla assay Or a plasma PCR

disease; definitions, clinical severity scores, and newsuch as AMPLICOR for the purpose of administering rapid

syndromes. Scand. J. Infect. Dis., Suppl. 99, 87‑89.
1 1. Boivin, G., Handfield, J., Toma, E., Murray, G., Lalonde,

preemptive treatment.

The AMPLICOR qualitative and quantitative assays, as

R･, Tevere, V･ J･, Sun, R･and Berg?ron, M･ (1998): Evalua一well as the antigenemia assay were considered to be useful

血ofAMPLICOR cytomegalovl㌣S test With specimens

for the diagnosis and monitoring of CMV disease in immun0‑
compromised patients, including those with BMT, KT, andAIDS.

from human immunodeficiency vlruS‑infected Subj ects.
J. Clin. Microbio1., 36, 2509‑25 13.
12. Tong, C., Cuevas, L., Williams, H. and Bakran,A. (1998):
Use of laboratory assay to predict cytomegalovirus disease
in renal transplant reclpients. ∫. Clin. Microbio1., 36,

REFERENCES

268ト2685.

I. Gondo, H., Minematsu, T., Harada, M., Akashi, K.,

13. Pellegrin, I., Gamgue, I., Binquet, C., Chene, G., Neau,
D･, Bonot, P･, Bonnet, F･, Fleury, H･ and Pellegrin, J. L.

Hayashi, S., Taniguchi, S., Yamasaki, K., Shibuya, T.,
Takamatsu, Y., Teshima, T., Eto, T., Nagafuji, K., Mizun0,(1999): Evaluati?n of new quantitatiye assay for

S., Hosoda, K., Mori, R., Minamishima, Y. and Niho, Y.

diagnosis and monltOrlng Of cytomegalovlmS disease in( 1 994): CytomegaloviruS (CMV) antigenemia for rapid
diagnosis and monitoring of CMV‑ associated disease

human immunodeficiency virus‑positive patients. J. Clin.

Microbio1., 37, 3 124‑3 132.
14･ Aitken, C･, Barrett‑Muir, W., Millar, C., Templeton, K.,Thomas, J" Sheridan, F., Jeffries, D., Yaqoob, M. andafter bone marrow transplantation. Brit. J. Haemato1.,
86, 130‑137.

2･ Freymuth, F., Germetay, E., Petitjean, J., Eugene, G., Ligny,

Breuer･ J･ (1999): Use ofmolecular assays in diagn?sis

B. H., Rykelynck, J. P.,Legoff, C., Hazera, P. and Bazin, C.

and monitorlng Of cytomegalovirus disease fbllowlng( 1 994): Comparison of nested PCR for detection ofDNA

renal transplantation. ∫. Clin. Microbio1., 37, 2804‑2807.

in plasma with pp65 16ukocyte antigenemia procedure

15･ Boeckh, M., Gooley, T. A., Myerson, D., Cunningham,
T･, Schoch, G･ and Bowden, R･ (1996): Cytomegalovints

for diagnosis of human cytomegalovirus infection. J.

Clin. Microbio1., 32, 1614‑1618.
3. Barrett‑Muir, W., Aitken, C., Temleton, K., RaRery, M.,

pp65 antigenemia‑guided early treatment with ganciclovlr

Kelsey, S. and Breuer, J. (1 998): Evaluation of the murex

transplantation: a randomized double‑blind study. Blood,

versus ganciclovir at engraRment a洗er allogeneic marrowhybrid capture cytomegelovirus DNA assay versus

88, 4063‑4071.

16. Einsele, H., Ehninger, G., Hebart, H., Wittkowski, K.,
Schuler, U･, Jahn, G., Mackes, P., Herter, M., Klingebiel,plasma PCR and shell vial assay fわr diagnosis of humancytomegalovirus viremia in immunocompromisedT., Loemer, J., Wagner, S. and Mueller, C. (1995):

patients. ∫. Clin. Microbio1., 36, 2554‑2556.

4. Dykewitz, C. A., Jaffe, H. W., Koplan, J. E. (2000):

Polymerase chain reaction monitoring reduces the

Guidelines for preventing OPPOrtunistic infections amongincidence of cytomegalovims disease and the duration

hematopoletic stem cell transplant reclplentS. Morbid.

and side effect of antiviral therapy after bone marrow

Mortal. Wkly. Rep., 49, RR No.10,ト14.

transplantation. Blood, 86, 28 1 5‑2820.

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